1. Biosimilar : Quality Comparability Case Wisit Tangkeangsirisin, PhD And Silpakorn Team

2. ALL Case Brief (1) The case study examples describe the comparative analysis of a biosimilar mAb candidate with its RBP. The development of a biosimilar mAb can easily require the consideration of 50 or more quality attributes. A subset of 17 illustrative quality attributes was chosen.

3. Case Brief (2) Table 1 summarizes these quality attributes and the methods used for their determination. They cover of attributes of glycosylation, physico-chemical modifications and different biological functions.

4. 4

5. The following are known to modulate the glycoform: (1) Gnt I−/− Lec1 Mutant, (2) β(1.4)-GalT +, (3) α(2,3) Sialyltransferase +, (4) CMP-sialic acid transporter +, (5) GnT III+ Mutant, (6) Fut8−/− Mutant, (7) antisense RNA sialidase, (8) Sialidase activity, (9) low glucose/gln ( 100 mmHg), (16) DMSO, (17) glycerol, (18) low temperature (30–32◦C), (19) low cell viability, (20) shear stress, and (21) N-acetylmannosamine

6. Functions of mAbs Case I and II Case III

7. Target cell Complement ADCC Antibodies – Nature’s pro-drugs

9. N-linked glycosylation Representative structures of the three classes of N-linked glycans showing both monosaccharide composition and bond linkage information (note: the number of potential structures is too large to represent symbolically, and the glycans shown are merely representative of the types of structures within each category) Optimal and consistent protein glycosylation in mammalian cell culture

10. Non-human glycan structures in available glycoprotein expression systems

12. Case I

13. Deamidation, oxidation, formation of dimers and higher aggregates are common degradation sensitive marker for these types of degradation events. Dimers and aggregates are considered to have a potential to increase the risk of immunogenicity and are typically rated as critical quality attributes which need to be tightly controlled.

14. the primary structure is identical and higher order structure is considered to be the same host cell proteins, DNA, or toxic process reagents were confirmed to be compliant quality attribute ranges were generated from 10 RBP and 10 SBP candidate batches

15. Case I and Case II Compare SBP 1 or 2 vs. RBP A RBP A is a monoclonal IgG1 type antibody used for treatment in oncology and autoimmune disorders. The mAb is designed to bind a cell surface receptor and subsequently triggers Fc mediated cell killing via CDC, ADCC and apoptosis.

16. Case III Compare SBP 3 vs. RBP B RBP B is a monoclonal IgG1 type antibody used for treatment of autoimmune disorders. The mAb is designed to bind a soluble target, not found on cell surfaces. The mode of action consists of the binding and subsequent clearance of the soluble target. No Fc functionality has been observed as relevant for the in vivo activity.

17. Each sub Group Determine which quality attributes are critical. Rate each quality attribute as positive, neutral or negative when compared to RBP. Discuss whether the candidate could be qualified as SBPs at the quality level How remaining questions could be addressed by further studies or by the nonclinical and clinical parts of the development program?

18. Case I Discussion and Wrap up Class Presentation Case I, II, III Case II Discussion and Wrap up Case III Discussion and Wrap up IAIBICIIAIIBIIC III A III B IIIC 20 Min for group discussion 10 Min for case consensus 15 Min for case presentation

19. Have fun