Biosimilars of Monoclonal Antibodies E-Book

• Herausgeber: John Wiley & Sons
• Kategorie: Wissenschaft und neue Technologien
• Sprache: Englisch

Seitenzahl: 1278

Veröffentlichungsjahr: 2016

Beschreibung

Addressing a significant need by describing the science and process involved to develop biosimilars of monoclonal antibody (mAb) drugs, this book covers all aspects of biosimilar development: preclinical, clinical, regulatory, manufacturing. * Guides readers through the complex landscape involved with developing biosimilar versions of monoclonal antibody (mAb) drugs * Features flow charts, tables, and figures that clearly illustrate processes and makes the book comprehensible and accessible * Includes a review of FDA-approved mAb drugs as a quick reference to facts and useful information * Examines new technologies and strategies for improving biosimilar mAbs

Leseprobe

Table of Contents

Cover

Title Page

Notes on Contributors

Preface

1 The History of Therapeutic Monoclonal Antibodies

1.1 Summary

1.2 Introduction

1.3 New Markets for Old Antibodies, Old Markets for New Antibodies

1.4 Antibody Engineering: A New Approach to the Treatment of Disease

1.5 Fully Human Antibodies, What Else?

1.6 Antibody Design

1.7 Antibody Production

1.8 Recombinant Antibodies: No Limits…

Acknowledgments

References

2 Structure, Classification, and Naming of Therapeutic Monoclonal Antibodies

2.1 Summary

2.2 Introduction

2.3 Antibody Structure

2.4 Classification of Antibodies

2.5 IgG Subtype

2.6 Nomenclature of Therapeutic mAbs

2.7 List of Therapeutic mAbs on Market or in Review in the European Union and the United States

References

3 Mechanism of Action for Therapeutic Antibodies

3.1 Introduction

3.2 Blockade of Ligand–Receptor Interaction

3.3 Target Depletion via ADCC and CDC

3.4 Engaging Cytotoxic T Cell Through the Use of Bispecific Abs

3.5 Receptor Downregulation by Enhanced Internalization and Degradation

3.6 Targeted Drug Delivery

3.7 Summary

References

4 Therapeutic Monoclonal Antibodies and Their Targets

4.1 Summary

4.2 Introduction

4.3 Monoclonal Antibody Therapies for Infectious Diseases

4.4 Monoclonal Antibody Therapies for Autoimmune Diseases

4.5 Therapeutic Monoclonal Antibodies Against Neoplastic Diseases

4.6 Conclusion

References

5 Antibody Posttranslational Modifications

5.1 Summary

5.2 Introduction

5.3 Overview of Co‐ and Posttranslational Modifications

5.4 Glycosylation

5.5 Glycation

5.6 IgG‐Fab Glycosylation

5.7 The Influence of Expression Platform on CTM/PTMs and Unintended Physicochemical Changes

5.8 Human Antibody Isotypes Other than IgG

5.9 Conclusion

References

6 The Pharmacology, Pharmacokinetics, and Pharmacodynamics of Antibodies

6.1 Summary

6.2 Introduction

6.3 Pharmacology of Anticancer MAbs

6.4 Antibody Pharmacokinetics

6.5 Pharmacodynamics

6.6 Conclusions

References

7 Monoclonal Antibodies

7.1 Summary

7.2 Introduction

7.3 Ado‐trastuzumab Emtansine (Anti‐HER2 Antibody Conjugated with Emtansine, Kadcyla)

7.4 Alemtuzumab (Campath, Campath‐1H)

7.5 Bevacizumab (Avastin)

7.6 Brentuximab Vedotin (Anti‐CD30 Antibody, Adcetris)

7.7 Cetuximab (Anti‐EGFR Antibody, Erbitux)

7.8 Denosumab (Anti‐RANKL Antibody, Xgeva™; Prolia™)

7.9 Eculizumab (Anti‐C5 Antibody, Soliris)

7.10 Ibritumomab Tiuxetan (Anti‐CD20 Antibody, Zevalin)

7.11 Ipilimumab (Anti‐CTLA‐4 Antibody, Yervoy)

7.12 Obinutuzumab (Gazyva)

7.13 Ofatumumab (Anti‐CD20 Antibody, Arzerra)

7.14 Panitumumab (Anti‐EGFR Antibody, Vectibix™)

7.15 Pembrolizumab (Keytruda)

7.16 Pertuzumab (Perjeta)

7.17 Ramucirumab (Cyramza)

7.18 Rituximab (Anti‐CD20 Antibody, Rituxan)

7.19 Tositumomab and Iodine I‐131 Tositumomab (Anti‐CD20 Antibody, Bexxar)

7.20 Trastuzumab (Anti‐HER2 Antibody, Herceptin)

References

8 Development of Biosimilar Rituximab and Clinical Experience

8.1 Summary

8.2 Introduction

8.3 Reditux Development Overview

8.4 Preclinical and Toxicology Studies

8.5 Clinical Evaluation

8.6 Conclusions

References

9 Monoclonal Antibodies for Infectious Diseases

9.1 Summary

9.2 Into the Future: Prophylaxis and Precision Medicine

9.3 Immune Therapy: A Noble Undertaking that Went to the Dogs

9.4 What’s Taking So Long?

9.5 Staphylococcus aureus: Still Public Enemy Number One?

9.6 Pseudomonas aeruginosa: The Bacterial Cockroach

9.7 Immune Evasion and Degree of Difficulty

9.8 Clostridium difficile: You Can’t Win for Losing

9.9 If Two Is Enough, Is Six Too Many? mAb Combos

9.10 Prophylaxis or Therapy? When You Come to a Fork in the Road, Take It

9.11 Influenza and Plan “B”

9.12 Safety: Human Enough for You?

9.13 Another Precinct Is Heard from Immunomodulatory Agents for the Treatment of Chronic Infections

9.14 Are We There Yet? Easy to Use, Fast Turnaround, Point‐of‐Care Diagnostics

9.15 Yeah but Aren’t These (Biologic) Drugs Going to Be Expensive?

References

10 Monoclonal Antibodies for Musculoskeletal, CNS, and Other Diseases

10.1 Summary

10.2 Natalizumab (Tysabri)

10.3 Eculizumab (Soliris)

10.4 Ranibizumab (Lucentis)

10.5 Denosumab (Prolia and Xgeva)

10.6 Antibody Therapies for Solid Organ Transplantation (Muromonab‐CD3 (Orthoclone OKT3), Basiliximab (Simulect), and Daclizumab (Zenapax))

10.7 Conclusion

References

11 Manufacture of Recombinant Therapeutic Proteins Using Chinese Hamster Ovary Cells in Large‐Scale Bioreactors

11.1 Summary

11.2 Introduction

11.3 Process and Cells: The Quasi‐species Concept Explains Individualized Development Needs

11.4 Choices for Manufacturing: Host Cells for Production and Suitable Selection Systems

11.5 Methods for Rapid Generation of High‐Producing Cell Lines

11.6 Silencing: Stability of Expression, Facilitators for High‐Level Productivity

11.7 High‐Throughput Bioprocess Development

11.8 Disposable Bioreactors

11.9 Nonclonal Expression Technologies for Fast Production and Assessment of Expression Potential and Quality

11.10 Conclusions

Conflict of Interest

References

12 Process Development

12.1 Summary

12.2 Introduction

12.3 Protein A and Protein G Batch Affinity Chromatography

12.4 Alternatives to Protein A

12.5 Disposables and Continuous Downstream Processing

12.6 Conclusion

References

13 Biosimilars and Biobetters

13.1 Summary

13.2 Introduction

13.3 The Biosimilar Pipeline

13.4 Developing Countries Will Continue to Prefer Cheaper Biogenerics

13.5 Biosimilar Candidates in the Pipeline

13.6 Biosimilar Development by Country/Region

13.7 Biosimilars Impact on Biopharmaceutical Markets and the Industry

13.8 Marketing Biosimilars Will Be a Challenge

13.9 Biosimilar Manufacturing Will Be State of the Art

13.10 Biosimilars Will Increase Demand for Product Quality and Transparency

13.11 CMOs Benefit from Biosimilars

13.12 Conclusions

References

14 Cell Line and Cell Culture Development for Biosimilar Antibody‐Drug Manufacturing

14.1 Summary

14.2 Mammalian Cell Line Development

14.3 Cell Culture Process Development

14.4 Future Trends

References

15 Product Analysis of Biosimilar Antibodies

15.1 Summary

15.2 Introduction

15.3 Identity

15.4 Purity and Impurities

15.5 Stability

15.6 Quantity—Concentration Measurement

15.7 Biological Activity—Functional Bioassays

15.8 Efficacy and Safety: Animal Studies for Antibody‐Drug Efficacy, PK/PD, and Toxicity

References

16 Bioanalytical Development

16.1 Summary

16.2 Introduction

16.3 Pharmacodynamics Characterization

16.4 Pharmacokinetic Assessment

16.5 Immunogenicity Assessment

16.6 Conclusion

References

17 Preclinical and Clinical Development of Biosimilar Antibodies

17.1 Summary

17.2 Introduction

17.3 Quality and Preclinical Development of Biosimilar Monoclonal Antibodies

17.4 Extrapolation of Indications

17.5 Clinical Development of Biosimilars of Monoclonal Antibodies

17.6 Ongoing Trials of Candidate Biosimilars of Monoclonal Antibodies

17.7 Conclusion

References

18 Regulatory Issues

18.1 Summary

18.2 Introduction

18.3 Existing Regulatory Pathways

18.4 Challenges

18.5 Conclusion

References

19 Legal Considerations

19.1 Summary

19.2 Overview of the Biologics Price Competition and Innovation Act of 2009 (“BPCIA”)

19.3 Patent Litigation and the BPCIA

19.4 Patenting Your Biosimilar

19.5 Conclusion

20 ADCC Enhancement Technologies for Next‐Generation Therapeutic Antibodies

20.1 Summary

20.2 Introduction

20.3 Activation of ADCC Functions

20.4 ADCC Enhancement through Glycol‐Engineering Technologies

20.5 Major ADCC Enhancement through Glycol‐Engineering Technologies

20.6 ADCC Enhancement through Fc Mutagenesis

20.7 Major ADCC Enhancement Fc Mutagenesis Technologies

20.8 Conclusion

References

21 Antibody Half‐Life

21.1 Summary

21.2 Introduction

21.3 The IgG Molecule as a Therapeutic Entity

21.4 FcRn and Antibody Half‐Life

21.5 Optimizing Antibody Fragments’ Half‐Life

21.6 Albumin Fusions for Half‐Life Extension

21.7 Mice as Models for Human Disease

21.8 Half‐Life Engineering: Present and Future

21.9 A Bright Future for Biosimilars, Biobetters, and Improved Half‐Life Modifications

References

22 Technologies for Antibody‐Drug Conjugation

22.1 Summary

22.2 The Importance of Therapeutic Index

22.3 ADC Construction: Building from the Protein Out

22.4 Conjugation Sites and Heterogeneity

22.5 Installation of Conjugation Sites

22.6 Bioconjugation Reactions

22.7 Linking Antibodies and Payloads

22.8 Conclusion

References

Index

End User License Agreement

List of Tables

Chapter 02

Table 2.1 Overview of antibody classes and subclasses.

Table 2.2 Physiochemical properties of human IgG subclasses.

Table 2.3 Biological properties of human IgG subclasses.

Table 2.4 Commonly used source and target substems in generic names of mAbs.

Table 2.5 Therapeutic mAbs approved or in review in the European Union or United States.

Chapter 03

Table 3.1 Antibodies against soluble ligands.

Table 3.2 Properties of TNF mAbs.

Table 3.3 MAbs to cell surface receptors.

Table 3.4 Armed mAbs.

Chapter 04

Table 4.1 An overview of the therapeutic goals of mAbs.

Table 4.2 Mechanisms for unresponsiveness to mAb therapy.

Table 4.3 Examples of mAb targets for infectious diseases.

Table 4.4 Examples of mAb targets for autoimmune diseases.

Table 4.5 Examples of mAbs for cancer treatment.

Chapter 06

Table 6.1 Pharmacokinetic properties of anticancer antibodies.

Table 6.2 PD models of anticancer antibodies.

Chapter 07

Table 7.1 Duration of disease‐free survival in patients with adjuvant treatment of breast cancer (Studies 1 and 2) [40].

Table 7.2 Duration of disease‐free survival in patients with adjuvant treatment of breast cancer (Study 4) [40].

Chapter 08

Table 8.1 Physicochemical characterization tests for Reditux and MabThera comparisons.

Table 8.2 Pharmacokinetic parameters for Reditux.

Table 8.3 Pharmacokinetic parameters of Reditux and MabThera.

Chapter 10

Table 10.1 Fact sheet of the monoclonal antibodies mentioned in this chapter.

Table 10.2 Variable domain sequences of listed antibody drugs (CDRs are in Gray).

Chapter 13

Table 13.1 Some leading reference products and the number of biosimilars targeting these products.

Table 13.2 Leading companies (≥8 biosimilars in their pipeline), location, and number of biosimilars.

Chapter 16

Table 16.1 Chromatographic assays versus immunoassays.

Table 16.2 Analytical comparability assessment.

Table 16.3 Parallelism testing.

Table 16.4 Parallelism acceptance criteria.

Chapter 17

Table 17.1 Required comparability testing for biosimilar products.

Chapter 18

Table 18.1 Glossary of terms.

Chapter 19

Table 19.1 Exclusivity periods for various biological products under 351(k) of the PHS Act, as amended by the BPCIA.

Table 19.2 First‐round procedure for developing the list and description of patents (see Sections 19.3.1, 19.3.3, and 19.3.4).

Chapter 21

Table 21.1 Examples of engineered mouse and human Fc regions and their properties.

Table 21.2 SPR affinity measures of antibody variants and combinations at steady state on immobilized recombinant human FcRn of IgG‐WT and variants produced in YB2/0 cells.

Table 21.3 Approaches for extending IgG half‐life through modification of the FcRn.

Table 21.4 Outlook for therapeutic applications of half‐life modifications.

List of Illustrations

Chapter 02

Figure 2.1 Classification of therapeutic mAb according to its increasing content of human sequence.

Figure 2.2 General structure of antibody.

Figure 2.3 Five different types of mammalian Igs.

Figure 2.4 Population distribution of five types of antibodies in blood (%).

Figure 2.5 Subtypes of IgG human antibodies.

Chapter 05

Figure 5.1 Domain structure of IgG.

Figure 5.2 Representative IgG‐Fc complex diantennary oligosaccharide. The “core” heptasaccharide has the composition: (GlcNAc)2Man3(GlcNAc)2, in blue [69, 72–75].

Figure 5.3 Amino acid residues of the C

H

2 domain contributing interactions with the sugar residues: single‐letter code.

Chapter 07

Figure 7.1 Nomenclature of different types of monoclonal antibodies. Comparison of monoclonal antibodies (dark brown, human; light blue, nonhuman or mouse). Top row: murine mAbs (name ends in “‐omab”); ibritumomab tiuxetan. Top row: chimeric mAbs (name ends in “‐ximab”); example: rituximab. Bottom row: humanized mAbs (name ends in “‐zumab”); example: trastuzumab. Bottom row: chimeric/humanized mAbs (name ends in “‐xizumab”); non are FDA approved. Bottom row: human mAbs (name ends in “‐umab”); example: panitumumab. Note: Public domain image from Wikimedia Commons. Accessed on October 18, 2016, from https://en.wikipedia.org/wiki/Nomenclature_of_monoclonal_antibodies.

Chapter 08

Figure 8.1 Structural similarity of Dr. Reddy’s rituximab and originator product. Similar identity in amino acid sequence (a), secondary (b), and tertiary structure (c) for Reditux and originator products. Spectral profiles in red indicate method/batch to batch variability determined from Innovator product sourced in the European Union and the United States.

Figure 8.2 Functional similarity of Dr. Reddy’s rituximab and originator product. Cell‐based cytotoxicity induced by complement establishes CDC, cell‐based cytotoxicity induced by PBMCs establishes ADCC, and cell‐based assay for detection of annexin V‐positive cells establishes apoptosis. Assays used for binding comparisons include cell‐based competitive binding FACS assay for CD20 ligand binding; ELISA‐based binding assay for C1q, FcγRIIa, and FcγRIIb binding; and binding constant determination by surface plasmon resonance for FcγRI, FcγRIIIa, and FcRn binding. The figure shows CDC (a) and apoptosis (b) assays and the FcγRIIIa binding sensorgrams (c).

Figure 8.3 Five‐year progression‐free survival (months) for patients receiving Reditux or MabThera. (a) The 5‐year progression‐free survival (months) for all patients treated with cyclophosphamide, doxorubicin, vincristine, and prednisolone with rituximab. (b) Five‐year progression‐free survival (months) for patients receiving MabThera alone or Reditux alone in all cycles of cyclophosphamide, doxorubicin, vincristine, and prednisolone with rituximab.

Figure 8.4 Five‐year overall survival (months) for patients receiving Reditux or MabThera. (a) The 5‐year overall survival (months) of all patients treated with cyclophosphamide, doxorubicin, vincristine, and prednisolone with rituximab. (b) The 5‐year overall survival (months) of all patients treated with MabThera alone or Reditux alone in all cycles of cyclophosphamide, doxorubicin, vincristine, and prednisolone with rituximab.

Chapter 10

Figure 10.1 Mechanism of immune‐cell mediated inflammation and its inhibition by α4 antagonist natalizumab. Lymphocytes flowing in the blood vessels are induced via chemokines to slow down, roll along the blood vessel walls, and finally arrest their movement and attach to the endothelium through a complex sequence of events. The local release of chemokines at the site of inflammation causes upregulation of α4β1 and α4β7 on the circulating immune cells and adhesion molecules such as V‐CAM and/or MAdCAM, on the surface of proximal endothelial cells [6]. Increased adhesion molecule density facilitates contacts between circulating immune competent leukocytes and vascular endothelium. As blood flows, binding between P‐selectin and E‐selectin molecules and cell surface glycoprotein or α4 integrins and endothelial cell adhesion molecules provide the initial contacts needed to capture circulating leukocytes (tethering and rolling). The decreased velocity allows chemokines on the surface of vascular endothelium to bind G‐protein coupled receptor (GPCRs) on the leukocyte surface, transducing signals that further activate integrins (activation). The high avidity of activated α4 integrins to their respective ligands results in firm adhesion to the endothelium (arrest). Firm adhesion is required for the process of diapedesis, extravasation of leukocytes through the endothelium into the extracellular matrix (ECM), to occur. When α4 integrins are blocked by natalizumab, lymphocyte adhesion to the endothelial wall is disrupted as their interactions with I‐CAM and V‐CAM are impaired.

Figure 10.2 Mechanism of complement activation cascades and molecular action of eculizumab. The complement cascade can be activated via the classical, mannose‐binding lectin and alternative pathways. All pathways of complement activation converge at the cleavage of the terminal complement protein C5. The C5 convertase cleaves C5 to C5a and C5b. C5b binds C6, C7, and C8 to form C5b‐8 complex, which binds to several molecules of C9 to form the mature membrane attack complex (MAC) C5b–9. In PNH patients, the deficiencies of GPI‐anchored proteins CD55 and CD59, which are the consequences of mutant PIGA, lost protection of cells from attack by the alternative pathway of complement, thus resulting in intravascular hemolysis. Eculizumab binds to C5 and prevented its entry into the C5 convertase. This effect prevents C5 cleavage and formation of membrane attack complex, thus rescuing PNH patients from complement‐mediated intravascular hemolysis. In the case of aHUS, mutations are in the complement regulatory proteins (factor H, factor I, etc.) involved in the formation or regulation of the alternative C3 convertase complex, thus resulting in inefficient protection of the endothelium from complement attack.

Figure 10.3 Mechanism of VEGF signaling in angiogenesis and its inhibition by ranibizumab. VEGFR1 and VEGFR2 are two major VEGFRs on the endothelial cells involving in angiogenesis, and they are members of the receptor tyrosine kinase family. VEGF‐A binds to VEGFR1 and VEGFR2 and induces receptor phosphorylation and subsequent activation of signals downstream. VEGFR1 is required for the recruitment of hematopoietic precursors and migration of monocytes and macrophages, whereas VEGFR2 is essential for the function of vascular endothelial cells. Phosphorylation of VEGFR2 induces the MAPK/ERK signaling pathway, which contributes to cell proliferation and migration. Alternatively, VEGF‐A could also activate phosphatidylinositol‐3‐kinase (PI3K) pathway, which also contributes to cell migration. Downstream signaling of VEGFR2 is very complex; therefore only simplified illustrations are described here. Detail signaling pathways of VEGF receptor signaling in the control of vascular function could be found in Olsson’s review [31]. Ranibizumab binds to the receptor binding site of all isoforms of VEGF‐A. The binding of the ranibizumab to VEGF‐A prevents the interaction of VEGF‐A with its receptors on the surface of endothelial cells, thus inhibiting angiogenesis.

Figure 10.4 Mechanism of RANKL signaling in osteoclast activation and its inhibition by denosumab. Stromal/osteoblast cells can secrete RANKL. RANKL is type II transmembrane protein, which has a small N‐terminal intracellular domain, a transmembrane domain, and a C‐terminal extracellular domain consisting of a stalk and a receptor binding region to form trimmers [44]. RANKL exists in both membrane‐bound and soluble forms. Membrane‐bound RANKL exists on marrow stromal/osteoblastic cells, which modulates the production of OPG and RANKL to control the osteoclast activity. Soluble RANKL are generated from cleavage of the extracellular tail region or alternative splicing of membrane‐bound RANKL transcripts. The trimer form of soluble RANKL binds to three receptors (RANKs) on the membrane of osteoclast precursor, thus inducing receptor clustering and signaling. Activated RANK recruits TNFR‐associated factors (TRAFs) to receptor and activates intracellular signaling downstream, thus leading to osteoclast maturation and bone resorption. OPG is a novel TNFR family member that exists only as a secreted protein, and it is a natural inhibitor of RANKL. Denosumab binds to RANKL with high affinity (

K

d

of 3 pM) and inhibits its interaction with RANK.

Figure 10.5 Mechanism of T‐cell activations and sites of action of available monoclonal antibodies in allograft transplantation. Antigen‐presenting cells (APC) presents antigen through MHC, which bind to T‐cell receptor (TCR). Activation of TCR triggers various downstream, including IL‐2 production and CD25 expression. Secreted IL‐2 functions as autocrine and binds to “high‐affinity” IL‐2R, which includes all three subunits. The IL‐2 induced IL‐2R stimulation leads to the activation of Janus tyrosine kinases (JAKs) and the signal transducer and activator of transcription (STAT) molecules. Phosphorylated STAT molecules undergo tetramerization and translocate to the nucleus, where they can bind to promoter expression of key genes. PI3Ks are also involved and have key downstream signals, including activation of mTOR, a target of rapamycin. Sites of action of available monoclonal antibodies in allograft transplantation are shown here. Basiliximab and daclizumab bind with high affinity to the interleukin‐2 receptor (CD25) and prevent the formation of the IL‐2 binding site interrupting the cascade of cellular events such as T‐cell activation and expansion and cytokine release to promote B‐cell activation and effector functions. OKT3 is an immunoglobulin that targets the membrane glycoprotein CD3 epsilon chain on the surface of circulating human T cells, which is part of the T‐cell receptor complex. Thus, OKT3 blocks both the generation and function of cytotoxic T cells clearing them from the circulation. Also shown here are actions of a few small molecule drugs. Rapamycin inhibits cell cycle progression through its interaction with mTOR. Cyclosporin A and tacrolimus inhibit calcineurin, thereby inhibiting NFAT and IL‐2 synthesis.

Figure 10.6 The schematic structures of studied antibodies in this chapter. Monoclonal antibodies covered in this chapter illustrate the evolution of monoclonal antibody therapeutics. Muromonab (g) is a murine antibody. Basiliximab (f) is a human/murine chimera with whole variable domain from murine origin. Natalizumab (a), eculizumab (b), bevacizumab (c), and daclizumab (e) are humanized monoclonal antibodies with only murine CDRs. Ranibizumab (c) is a humanized antibody fragment (Fab) derived from bevacizumab. Denosumab (d) is a fully human monoclonal antibody.

Chapter 11

Figure 11.1 Derivation of spontaneously immortalized CHO cells from an ovary explant of a female, outbred Chinese hamster as referred to in the literature. Filled dots indicate events eluded to by traceable description or reference; wavy lines indicate nonreported details on subcultivation (passaging), culture media, and culture vessels.

Figure 11.2 History of CHO cell lines as obtained from the literature. Uninterrupted lines indicate known transfers of cell cultures to other labs. Wavy lines indicate uncertain and poorly recorded history, lacking details of culture conditions and number of subcultivations (passaging). The smaller arrows at boxed names of cells indicate probable, but unrecorded, transfer of cell cultures to other laboratories. Historic times indicated in the figure are approximate and deducted from various literature sources.

Chapter 12

Figure 12.1 Schematic of a protein A/G affinity chromatography purification. Clarified cell culture fluid is loaded onto a column containing protein A resin at a neutral pH (6–8). Protein A binds and captures the MAb, while HCP and other contaminants flow through the column. The column is washed, and relatively pure MAb is eluted from the column under low pH (2.5–4) conditions. Residual contaminants are removed in downstream polishing steps to yield a product of the required purity.

Figure 12.2 Schematic of PCC chromatography. Three (or more) columns are utilized in order to allow for continuous purification of the MAb from the feed. Cells 1 and 2: clarified cell culture fluid is loaded onto the first column with effluent directed to waste until breakthrough occurs. Effluent is redirected to column 2 to avoid product loss. Cells 3 and 4: column 1 is loaded to saturation, feed is loaded onto column 2 with effluent directed to waste, and column 1 is sequentially washed, eluted, regenerated, and re‐equilibrated. Cells 5 and 6: at the point of product breakthrough, effluent from column 2 is loaded onto column 3 to capture unbound MAb. Column 2 is then loaded to saturation. Cells 7–9: feed is loaded onto column 3 with effluent directed to waste, and column 2 is processed. At the point of breakthrough from column 3, effluent is redirected to column 1 to capture unbound MAb. Column 3 is loaded to saturation and the process is repeated.

Figure 12.3 Schematic of an SMB purification process. Countercurrent flow of resin and liquid is approximated by periodic rotation of multiple columns (numbered 1–4) in the direction opposite to fluid flow. This figure will describe the activities of one column, starting in zone III, as it makes a full rotation through the system. Cell 1: in zone III, a feed containing A (weakly adsorbed) and B (strongly adsorbed) flows through the column and some pure A elutes before B begins to co‐elute. This pure fraction of A is removed as the raffinate. Cell 2: the columns rotate and the unresolved mixture of A and B moves through the column while it remains in zone II. This leaves pure B near the column exit. Cell 3: the columns are rotated, desorbent is added to the column (now in zone I), and this pure fraction of B is eluted from the column. B is collected as the extract. Cell 4: the column, now free of all A and B, rotates into zone IV where it removes all A not collected through the raffinate port from the mobile phase. The columns then rotate and the process is repeated.

Figure 12.4 Schematic of CAC. Separation media (resin) is loaded into the annulus of two concentric cylinders and covered with glass beads. Clarified cell culture fluid is fed continuously onto the column at a fixed position from a nozzle. Eluent is added at a uniform flow rate to the entire annulus of the column and the column is rotated around its axis at constant angular velocity. Proteins move through the column at different velocities. The faster‐migrating proteins elute quickly (the column will have rotated a short distance) and the slower‐migrating proteins elute later (the column will have rotated further). Each protein elutes at a constant angle/position and may be collected continuously.

Chapter 13

Figure 13.1 US biosimilars launchable dates (by reference products,

n

 = 133, not products in the pipeline).

Figure 13.2 Biosimilars launchable dates by sum of current reference products sales ($millions).

Figure 13.3 Pipelines by phase of development. Note: All but ≥20 of the “marketed” biosimilars are biogenerics in international commerce.

Figure 13.4 Number of companies in involved in biosimilars by size. The previous figure shows the numbers of companies known to be active in biosimilars development by company size. The table in the following shows the number of biosimilars in development by leading companies, in terms of number of biosimilars having been reported in development.

Chapter 14

Figure 14.1 Expression systems for biologic products. Mammalian cells are the most popular systems for antibody production, bacterial cells are for antibody fragments or Fab production, yeast cells have heavy glycosylation issue, and insect cells produce proteins with heavy phosphorylation. Plant plan expression platforms for food proteins and animal systems for reagent production. Cell free systems are optimal for protein production toxic to the host cells.

Figure 14.2 Major CHO host cells. CHO‐K1 and DXB11 have closer lineage relation, but DG44 is a variant from the CHO prototype. All other cell lines are derived from the original three ancestral parent lines.

Figure 14.3 Production cell line development steps. From the gene transfection step to the final MCB/WCB point is dependent on the technologies available to each company. The timeline for each step is varied. A 6‐month timeline is currently feasible for most companies.

Figure 14.4 Four categories of cell line productivity. The stability falls into four categories: (a) productivity increases over time, (b) productivity stays no change, (c) productivity drops quickly after long period of generation time, and (d) productivity drops gradually over time. a and b are less common, while c and d are more common.

Figure 14.5 Cell line productivity. Numbers can reach over 10 g/l, due to high‐productivity cell lines, balanced media, and optimized bioreactor processes. Currently 3–5 g/l is standard in biopharma industry.

Figure 14.6 Bioprocess steps. Upstream (cell line and cell culture) and downstream (purification and formulation/fill–finish). Analytical processing is ideal for supporting both steps.

Figure 14.7 Process scale‐up. Manufacturing scale process development from multiwell scale to commercial production. Process scale‐down is for process characterization, validation, and manufacturing process troubleshooting and commercial process tech support; normally from manufacturing scale to benchtop bioreactor scales.

Chapter 15

Figure 15.1 Outlines of Product Analysis of Biosimilar Antibodies.

Figure 15.2 Molecular mass of the light chain of a biosimilar antibody (bottom) versus the originator antibody (top).

Figure 15.3 Biosimilar and originator products have the same tryptic peptide mapping and mass spectrometry sequences. Top panel: originator drug; bottom panel: biosimilar product. Some antibodies may contain an extra glycan at the Fab region in addition to the Fc site. The extra glycan has to be treated differently from the standard method due to difficulties in cleaving the glycans from the Fab region for analysis (see more details in the glycan analysis section).

Figure 15.4 A typical MS/MS spectrum of one of the tryptic peptides in Fc region. Peptide sequence (KGNLWDGHLVTLVSVV) is confirmed by b/y ions.

Figure 15.5 Far‐ and near‐UV spectra of three lots of biosimilar antibody and three lots of originator antibody.

Figure 15.6 Schematic representation of glycan structure in antibodies. G0F, G1F, G1F′, G2F, and G2F with sialic acid, G0, G1, G1′, and M5. Asn is the amino acid residue at the Fc region of monoclonal antibody that is linked to the glycans. Fuc, fucose; Gal, galactose; GlcNAc,

N

‐acetylglucosamine; Man, mannose; SA, sialic acid.

Figure 15.7 HILIC glycan profiles of monoclonal antibodies produced in CHO cells. The black line is from the originator; light gray and gray lines are from two biosimilar lots.

Figure 15.8 Deconvoluted LC–MS mass analysis of heavy chain of an IgG monoclonal antibody either from the originator (bottom panel) or from a biosimilar (low panel).

Figure 15.9 MS/MS spectrum of a disulfide peptide VTITCR = SGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTK. Top panel: from originator; bottom panel: from a biosimilar.

Figure 15.10 Confirmation of N‐terminal pyroglutamate from both biosimilar (Analyte) and originator mAb (Control).

Figure 15.11 Example of CE–SDS of Beckman PA 800 under nonreducing conditions.

Figure 15.12 The CEX chromatograms of a biosimilar antibody (left panel) and originator antibody sample (right panel) with carboxyl peptidase B (CpB) digestion (lower panel) or without CpB digestion (upper panel).

Figure 15.13 IEF gel analysis of antibody samples with CpB treatment (right panel) or without treatment (left panel). C, reference sample; S, biosimilar sample; pI, markers with different pI.

Chapter 16

Figure 16.1 Complexity in structure and function of monoclonal antibodies.

Chapter 17

Figure 17.1 Overall evidence of comparability required to support extrapolation to indications not clinically studied [13].

Chapter 20

Figure 20.1

Comparison of ADCC‐mediated tumor cell killing by enhanced and untreated anti‐HER2 antibody preparations.

The antibody was treated with freshly recruited, isolated human peripheral blood mononuclear cells possessing the V158/V158 genotype.

Figure 20.2

Dramatically improved tumor cell killing observed with human peripheral blood mononuclear cells possessing the 158F/158F genotype.

Note that minimal cell killing was observed with the wild‐type antibody.

Figure 20.3

Schematic illustrating human IgG1 bearing two N‐linked biantennary complex‐type oligosaccharides covalently attached at the conserved asparagine 297 (Asn297).

This site is located in the CH2 domain of the Fc region. Note composition containing a mannosyl‐chitobiose core structure in the presence or absence of a core fucose, a bisecting

N

‐acetylglucosamine (GlcNAc), and a terminal galactose and sialic acid.

Figure 20.4

Schematic outlining GlycoMab technology.

GlycoMab produces defucosylated mAbs in target cell lines through the introduction of a bisecting β‐1,4‐GlcNAc residue to N‐linked oligosaccharides. Subsequently, other central reactions including core fucosylation and conversion of the hybrid to complex glycans are blocked.

Chapter 21

Figure 21.1 Antibody fragments and albumin fusion proteins.

Chapter 22

Figure 22.1 Modular components of an antibody‐drug conjugate. A monoclonal antibody (mAb) (a) can be engineered (b) to contain reactive sites (c). These sites include amino acids (AAs) and posttranslational modifications (PTMs). Partner reagents are added to form a conjugate with the reactive site (c). A linker (d) is attached to a payload (e), which can include a release trigger. Synthesis of an ADC involves addition of an elaborated drug–linker to an antibody containing a reactive site (f and g).

Figure 22.2 Proven technologies for antibody‐drug conjugation. The 58 unique ADCs that have advanced to clinical trials (at the time of writing) are summarized by technology used. Note that most ADCs employ more than one technology. (a) Conjugation technologies (Section 22.6.2.4). Predominantly lysine and cysteine conjugation are used, with two engineered cysteine ADCs entering trials recently. (b) Linker technologies (Section 22.7.4.4). Noncleavable linkers account for approximately 20% of ADCs, and cleavable constructs (6.2) somewhat evenly distributed the three trigger mechanisms—proteolysis, reduction, and pH. (c) Modular structural components of linkers (6.1) that impart solubility and stability listed by frequency of use. (d) Toxin payloads. The process of choosing a toxin is described in excellent detail in a recent review [15]. eCys, engineered cysteine; DM1,

N

2

′‐deacetyl‐

N

2

′‐(3‐mercapto‐1‐oxopropyl)maytansine; DM4,

N

2

′‐deacetyl‐

N

2

′‐(4‐mercapto‐4‐methyl‐1‐oxopentyl)maytansine; DS, disulfide; MMAE, monomethyl auristatin E; MMAF, monomethyl auristatin F; ND, not disclosed; PBD, pyrrolobenzodiazepine; SN‐38, the active metabolite of irinotecan.

Figure 22.3 Bioconjugation to native AAs. Lysine (1) and cysteine (6) are endogenous sites on mAbs that can react with partner reagents. Common reaction conditions for conjugate formation are shown, along with notable liabilities. NR, not reported.

Figure 22.4 Bioconjugation to PTMs. Antibodies can contain native and nonnative PTMs that react with partner reagents. Common reaction conditions for conjugate formation are shown, along with notable liabilities. DHA, dehydroascorbic acid; DTT,

D

,

L

‐dithiothreitol; TCEP, tris(2‐carboxyethyl)phosphine.

Figure 22.5 Bioconjugation with enzymes on peptide‐tagged mAbs. Antibodies can contain AA sequences that are recognized by bond‐forming enzymes. Common reaction conditions for conjugate formation are shown. mTGase, microbial transglutaminase; PPTase, phosphopantetheinyl transferase.

Figure 22.6 Examples of secondary bioconjugation. Nonnatural functional groups are installed with heterobifunctional chemical reagents. Bioconjugation involves a bioorthogonal reaction joining payload and antibody. GalT, galactosyltransferase; SiaT, sialyltransferase.

Figure 22.7 Modular components used in linker preparation. Each component must be considered for stability and solubility. Synthetic organic chemistry enables preparation of drug–linker reagents for bioconjugation.

Figure 22.8 Triggers that can be included for linker cleavage. A variety of cellular components or environments are used to induce release of payload.

Figure 22.9 Self‐immolative spacers. These spacers enable automatic release of unmodified payload after a triggered event. Very few reliable technologies are available for this purpose.

Guide

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Biosimilars of Monoclonal Antibodies

A Practical Guide to Manufacturing, Preclinical, and Clinical Development

Edited by

Cheng Liu, Ph.D.K. John Morrow, Jr., Ph.D.

Copyright © 2017 by John Wiley & Sons, Inc. All rights reserved

Published by John Wiley & Sons, Inc., Hoboken, New Jersey

Published simultaneously in Canada

No part of this publication may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, scanning, or otherwise, except as permitted under Section 107 or 108 of the 1976 United States Copyright Act, without either the prior written permission of the Publisher, or authorization through payment of the appropriate per‐copy fee to the Copyright Clearance Center, Inc., 222 Rosewood Drive, Danvers, MA 01923, (978) 750‐8400, fax (978) 750‐4470, or on the web at www.copyright.com. Requests to the Publisher for permission should be addressed to the Permissions Department, John Wiley & Sons, Inc., 111 River Street, Hoboken, NJ 07030, (201) 748‐6011, fax (201) 748‐6008, or online at http://www.wiley.com/go/permissions.

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Library of Congress Cataloging‐in‐Publication Data:

Names: Liu, Cheng, 1966– editor. | Morrow, John, Jr. 1938– editor.Title: Biosimilars of monoclonal antibodies : a practical guide to manufacturing, preclinical, and clinical development / edited by Cheng Liu, K. John Morrow, Jr.Description: Hoboken, New Jersey : John Wiley & Sons, Inc., [2017] | Includes bibliographical references and index.Identifiers: LCCN 2016019958 (print) | LCCN 2016020707 (ebook) | ISBN 9781118662311 (cloth) | ISBN 9781118940624 (pdf) | ISBN 9781118940631 (epub)Subjects: | MESH: Antibodies, Monoclonal | Biosimilar PharmaceuticalsClassification: LCC QR186.7 (print) | LCC QR186.7 (ebook) | NLM QW 575.5.A6 | DDC 616.07/98–dc23

LC record available at https://lccn.loc.gov/2016019958

Cover image: Courtesy of the Editors

Notes on Contributors

K. Lance Anderson focuses his law practice at Dickinson Wright PLLC on intellectual property, business and information technology related to emerging business. A registered patent attorney, he has wide‐ranging experience in licensing and transactional matters, including negotiation and due diligence for licensing, mergers and acquisitions, asset transfers, and strategic planning. He often counsels on pharma and life science transactions, including research and development, in the developing anticancer and immunotherapeutic industries. His experience also includes patent prosecution, patent litigation, trade secrets, employment, corporate governance, electronic records management, and financial matters.

Lance earned his B.S. in Entomology and M.S. in Crop Science from Texas Tech University and his J.D. from the Texas Tech University School of Law.

Jonathan D. Ball, Ph.D., is a shareholder at Greenberg Traurig, LLP, with nearly 15 years of experience representing clients in intellectual property matters, with an emphasis on patent litigation and contentious Patent Office proceedings. He also has extensive experience in patent portfolio management, prosecution, and licensing and helps companies develop and implement worldwide IP strategies. He has experience in a variety of biopharmaceutical matters, including Hatch–Waxman litigation, pre‐ANDA diligence, and counseling on pharmaceutical patent life cycle management.

Jonathan earned his B.A. from Vassar College, his Ph.D. in Organic Chemistry from the University of North Carolina at Chapel Hill, and his J.D. from the University of Richmond School of Law.

Maurizio Chiriva‐Internati An author of 146 peer‐reviewed publications, Maurizio Chiriva‐Internati, Ph.D., D. Bsci., acts as the chief scientific officer at Kiromic, a biotechnology company specialized in immunotherapy, and serves as an associate professor of medicine and director of translational research at the Division of Hematology/Oncology in the Department of Internal Medicine at Texas Tech University Health Sciences Center in Lubbock, Texas. He received his Ph.D. in Immunology from the University of Nottingham and his doctorate in biology from the University of Milan. He has clinical and teaching experience in M.D., Ph.D., and M.S. school settings as well as experience as a reviewer for a variety of journals. He is an associate editor for the Journal of Translational Medicine, a section editor for the International Reviews of Immunology, and an associate editor member for both the Open Hematology Reviews and the Open Hematology Journal. For the past 14 years, he has investigated tumor antigens as therapeutic and diagnostic tools. His research is focused on developing new therapies for a variety of diseases, namely, cancers including multiple myeloma, ovarian, lung, prostate, melanoma, and breast. Using recombinant adeno‐associated viruses, he is developing vaccines that direct T‐cell immune responses toward unique tumor‐specific or tumor‐associated antigens.

Everardo Cobos, M.D., is president of Kiromic, LLC, and director of Hematologic Malignancies at Grace Health System in Lubbock, Texas. For the past 24 years, he served as a professor of medicine and oncology and chairman of the division of hematology/oncology at Texas Tech University Health Sciences Center (HSC) in Lubbock. He has also served as medical director of the Southwest Cancer Center at the University Medical Center (UMC) and as program director and founder of the Stem Cell Transplantation Program at TTUHSC/UMC. He is board certified in internal medicine, hematology, and oncology and is an expert in the treatment of hematologic malignancies, stem cell transplantation, and thrombosis and hemostasis.

Dr. Jose A. Figueroa is currently director of Solid Malignancies in the Blood Disorders and Cancer Therapeutics Center at Grace Health System in Lubbock, Texas, and vice president and chief medical officer of Kiromic, LLC, a clinically oriented biotechnology company. He earned his undergraduate and medical degrees at the University of Puerto Rico and completed his fellowship in medical oncology, hematology, and translational research at the University of Texas Health Science Center in San Antonio in 1993. He is board certified in medical oncology. He has been a recipient of the Young Oncologist Award from the American Radium Society and the American Cancer Society Research Grant and has served as medical director and director of the clinical research program of the Southwest Cancer Center/University Medical Center at Texas Tech University Health Sciences Center, School of Medicine. He is the author of many scientific abstracts, peer‐reviewed publications, a book chapter on breast cancer, poetry, and a fictional novel.

João Eurico Fonseca is head of the Rheumatology Research Unit at the Instituto de Medicina Molecular and professor of rheumatology and biomedical engineering, both part of the Faculty of Medicine, University of Lisbon, Portugal. He is also head of the Ambulatory Unit of the Rheumatology Department of the Lisbon Academic Medical Centre. He completed his M.D. and Ph.D. in Rheumatology at the University of Lisbon and has obtained a qualification in health management from the AESE School of Management and Business in Lisbon. His research focuses on bone and inflammatory joint diseases. He is also the president of the Portuguese Society of Rheumatology and a member of the editorial board of several medical journals. He has authored more than 150 scientific papers.

João Gonçalves is head of the Biopharmaceutical and Molecular Biotechnology Unit at the Institute of Innovative Medicines and professor of immunology and biotechnology, both part of the Faculty of Pharmacy, University of Lisbon, Portugal. He is also head of Antibody Engineering Laboratory at iMed—Faculdade Farmacia Universidade Lisboa. He has a pharmaceutical degree and Ph.D. in Infectious Diseases/Immunology at Harvard University and University of Lisbon and has postdoctoral experience at Centocor, Genentech, and Scripps Research Institute, La Jolla. His research focuses on therapeutic antibody discovery and development ranging from autoimmune diseases to oncology. He is a member of the Portuguese Pharmacopoeia Commission and Portuguese Commission of Medicines Evaluation. He is also a member of the editorial board of several scientific journals. He has authored more than 80 scientific papers.

Patrick G. Holder is a native of Cleveland Heights, OH. He received his Ph.D. in Chemical Biology in the lab of Matthew Francis at UC Berkeley. While there, he used bioconjugation to integrate optoelectronic materials into a scaffold of self‐assembling viral proteins, including single‐walled carbon nanotubes, arrays of phthalocyanines, and thermostable virus‐templated polymers. As an NIH postdoctoral fellow in the lab of Daniel Nocera at MIT, in a collaboration with JoAnne Stubbe, he leveraged site‐specific bioconjugation of phototriggers to perform time‐resolved spectroscopy of radical transport in the enzyme ribonucleotide reductase. In 2012, his work was awarded the Energy and Environmental Science Prize from the Royal Society of Chemistry. He graduated with an A.B. in Chemistry from Columbia University, where he received the William H. Reinmuth Chemistry Scholarship. Formerly at Redwood Bioscience and currently at Bristol‐Myers Squibb, he is helping advance antibody‐drug conjugates from the benchtop to the clinic.

Dr. Nattamol Hosiriluck is currently a second‐year resident in the Department of Internal Medicine of Texas Tech University Health Sciences Center in Lubbock, Texas. She graduated from Phramongkutkloa College of Medicine in Bangkok, Thailand, in 2012. Since then her interest has been on hematology and oncology, and she is planning to pursue her fellowship in this area.

Clarinda Islam is currently the director of Quality Assurance and Regulatory Affairs for Somru BioScience, Inc., an emerging biotechnology company based in Charlottetown, PE, Canada. In this role, she is responsible for the quality management, project management, process improvement, product development life cycle methodologies, and quality management systems. Previously she served as a senior group leader for the Immunochemistry Department at PPD (VA, USA) where she was responsible for leading a team of scientists dedicated to supporting biopharmaceutical development. Prior to joining PPD, she has held positions of increasing responsibility in the Bioanalytical Group at Covance, Inc. She has over 14 years of experience in the development of biologics in a highly regulated environment under FDA and other global regulatory guidelines.

Rafiq Islam is the senior director of Bioanalytical Services at Celerion, Inc. (NE, USA). In his current role, he is responsible for the scientific and operational leadership of both small‐ and large‐molecule bioanalysis, which includes the development, validation, and execution of sample analysis to support pharmacokinetic, immunogenicity, bioequivalence, and biosimilar studies. Previously, he served as the scientific director for Biopharma Services at EMD Millipore. He held similar positions as the head of a bioanalytical department at Covance, Inc. (AZ, USA), and Huntingdon Life Sciences (NJ, USA). He also held several positions of increasing responsibility with CuraGen Corporation (New Haven, CT).

He has a B.S. degree in biology and M.Sc. in data mining. He has 15 years of industry experience. He is an active member of the American Association of Pharmaceutical Scientists (AAPS) and Global CRO Council (GCC).

Roy Jefferis After receiving his B.Sc. and Ph.D. in Chemistry, Jefferis moved to the Medical School (Birmingham) to study the structure and function of antibody molecules. A particular interest has been the influence of glycosylation on the expression of biologic activities, in vitro and ex vivo. Extension of these findings to glycoengineering and protein engineering of recombinant antibody therapeutics resulted in interactions with the biopharmaceutical industry. A current focus is the potential for immunogenicity of immune complexes, in contrast to aggregated forms of IgG. He is the author of more than 280 publications, with 207 of these referenced in PubMed. He is a member of the Royal College of Physicians (MRCP) and a fellow of the Royal College of Pathologists (FRCPath).

Maria de Jesus Trained as an environmental engineer in Portugal, De Jesus completed her Ph.D. in Plant Cell Culture Technology at the Swiss Federal Institute of Technology in Lausanne, Switzerland (EPFL). She joined the Laboratory of Cellular Biotechnology at the EPFL as a postdoctoral researcher and led the development of several projects as scientist and senior scientist, directed toward the establishment of manufacturing processes for recombinant proteins from CHO cells. In 2002 she initiated a self‐funded, privately owned company, ExcellGene SA, acting in the role of founder and first employee. In subsequent years she established a laboratory and administrative unit of this company while simultaneously assuring the profitability and growth of the company as a CRO and CMO in the field of protein manufacture and process development for high‐value therapeutic products from CHO and other cell lines. Currently she is chief operations officer, a member of the board of directors, and a significant shareholder of the company.

She has published over 40 peer‐reviewed papers in the field and has lectured on process development and scale‐up with animal cells in bioreactors. She is inventor of a small‐scale bioreactor system, based on orbitally shaken tubes which are used in high‐throughput approaches for suspension culture process development.

Dr. Weidong Jiang is a cofounder of Henlius Biopharmaceuticals, Inc. in United States. He is also senior vice president and chief scientific officer of Shanghai Henlius Biotech Co., Ltd., a joint venture formed by Henlius and Fosun Pharma in 2009 to develop antibody therapeutics. He has more than 20 years of working experience in many biotech companies, including Catalyst Biosciences, Inc.; Vasgene Therapeutics, Inc.; Applied Molecular Evolution, Inc. (subsidiary of Eli Lilly); Microcide; and ChemGenics (acquired by Millennium). He has expertise in the field of drug screening and development, particularly in research and development of antibody therapeutics and other protein biologics. He holds a Ph.D. degree from Giessen University in Germany; a master’s degree from Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences; and a bachelor’s degree from Zhejiang University.

Cheng Liu is the founder and CEO of Eureka Therapeutics, a California company dedicated to the development of innovative cancer immunotherapy. He worked at Chiron (now Novartis), where he championed the development of first‐in‐class therapeutic antibody against CSF‐1, a key immune regulator, for cancer metastasis treatment. He has multiple issued US and international patents and scientific publications, including in journals of Science and Nature, in the area of therapeutic antibody discovery and engineering. In 2007, he was awarded Special US Congressional Recognition for his contributions to improving human health. He received his B.S. in Cell Biology and Genetics from Beijing University and a Ph.D. in Molecular Cell Biology from the University of California, Berkeley.

Dr. Margaret Liu is an assistant professor in the Department of Chemical and Biological Engineering at the University of Alabama. Her research areas include mammalian cell engineering, microorganism metabolic engineering and systems biology, and bioreactor process development. Specifically, her research projects focus on (i) improving the production and quality of biopharmaceuticals (e.g., biosimilars and other recombinant proteins) by host cell engineering and integrated process development via omics technologies, (ii) developing novel therapy to treat cancer and heart diseases, and (iii) producing next‐generation bioenergy (e.g., biobutanol) by metabolic cell‐process engineering (MCPE) using systems biology. She had worked in biopharmaceutical and biotechnology industries for six and half years before joining academia research. She was a senior scientist and team leader of Life Technologies Corporation (LTC) and a scientist of Lonza and Merck KGaA (EMD biopharmaceuticals). She has years of diverse industrial experiences in mammalian cell culture, biopharmaceutical production cell line development, omics technology, and therapeutic protein (including both innovator biologics and biosimilars) production process development in pharmaceutical and biotech companies. She accomplished her Ph.D. research under the instruction of Prof. S.T. Yang in the Department of Chemical and Biomolecular Engineering at the Ohio State University, United States. Her Ph.D. thesis work dealt with bioprocessing of high‐value and biofuel products from metabolically engineered microbial mutants using a novel bioreactor production bioprocess. She earned her master’s degree in bioengineering from Tianjin University and bachelor’s degree in chemical engineering (major) and computer science (minor) from Shandong University, China.

Meimei Liu is a research scientist at Thermo Fisher Scientific, Inc. in Grand Island, NY. She obtained her B.S. and M.S. in Chemical Engineering from Tianjin University, China, and Ph.D. in Chemical and Biomolecular Engineering from the Ohio State University. While working at Tianjin University, she studied drug delivery and biopolymer development. While working at the Ohio State University, she investigated cell culture, cell therapy, and bioreactor process development. At Thermo Fisher Scientific, Inc., she works on cell culture and media development.

Dr. Scott Liu is a molecular biologist by training and holds a Ph.D. degree from Purdue University. As the founder, president, and CEO of Henlius Biopharmaceuticals, Inc. and Shanghai Henlius Biotech Co., Ltd., he takes overall responsibility for strategic planning and implementation, as well as operations management. Benefiting from Scott’s vision in achieving quality at reasonable cost, Shanghai Henlius Biotech Co., Ltd. has adopted innovative technologies in mAb development and manufacturing and, in doing so, significantly reduced the manufacturing costs while maintaining the quality of the products. Before founding his own companies, Scott held key technical and/or leadership positions in several biopharmaceutical companies, including Amgen, Bristol‐Myers Squibb, United Biomedicals (UBI), Tanox, and Maxygen. He was the director of Quality Analytical Labs of Amgen Fremont and took responsibility for quality control operations for late‐phase development and cGMP commercial manufacturing of Vectibix. Prior to Amgen, Scott held the post of associate director of Biologics QC in BMS Syracuse, where he was in charge of QC operations for late‐phase development and commercial manufacturing of Orencia.

James D. Marks Dr. Marks is an internationally recognized pioneer in the field of antibody engineering and an elected member of the Institute of Medicine of the National Academies of Sciences. He has invented broadly applicable methodologies for the generation and optimization of human monoclonal antibodies. The Marks Lab at UCSF has been involved in the use of diversity libraries and display technologies to generate monoclonal antibodies (mAbs) since the earliest days of the technologies. As a graduate student in Greg Winter’s Lab, he developed the first universal PCR primers which could amplify and isolate human V‐gene repertoires and was the first to show that human mAbs could be directly isolated from nonimmune (naïve) phage antibody libraries. He also was the first to demonstrate the use of phage display for in vitro