Date recorded: May 15 2018

Duration: 32 minutes 06 seconds

The modular nature of antibodies allows the design of antibody-related formats with tailored characteristics. The antibody’s Fc part, encompassing the effector functions, represents an attractive scaffold for engineering therapeutic molecules as demonstrated by the generation of Fcabs (Fc domain with antigen-binding sites). By engineering the C-terminal loops (AB, CD and EF loop) in the CH3 domains, two antigen binding sites can be inserted in close proximity.

The study presented in this webinar deals with the determination of the first X-ray structures of three Fcabs that differ in loop design, specificity, affinity and thermal stability and reveals their overall structural integrity and native fold.

Furthermore, by investigation of the interaction of these Fcabs with their respective antigens (HER2 or VEGF) using a set of complementary techniques (X-ray crystallography, size exclusion chromatography combined with multi-angle light scattering [SEC-MALS], isothermal titration calorimetry [ITC], fluorescence correlation spectroscopy [FCS]), insights into the binding modes and binding stoichiometries were gained.

Table of contents
1. Welcome
00:22
2. Elisabeth Lobner
00:11
3. Enlightening the Fcab-antigen interaction via X-ray crystallography & solution studies
01:08
4. Enlightening the Fcab-antigen interaction via X-ray crystallography & solution studies
00:35
5. Projects
00:38
6. Introduction: Modular Antibody Technology – How to get an Fcab?
00:07
7. IgG1-Fc binds them all
00:49
8. Modular Antibody Engineering
00:25
9. Modular Antibody Engineering
00:33
10. Fcabs under study
01:33
11. Fcabs under study
00:41
12. Project I: Fcab-HER2 interaction: A ménage à trois
00:00
13. X-ray structures of Fcabs
00:14
14. X-ray structures of Fcabs
00:53
15. X-ray structures of Fcabs
01:55
16. HER2
01:44
17. Targeting HER2
00:46
18. SEC-MALS
00:36
19. SEC-MALS
00:13
20. SEC-MALS
00:36
21. X-ray structure of Fcab‒HER2 complexes
00:46
22. X-ray structure of H10-03-6‒HER2 complex
00:35
23. ITC measurement of STAB19-HER2 interaction
00:23
24. Discrepancy in binding stoichiometries
00:07
25. Discrepancy in binding stoichiometries
00:47
26. FCS measurements of STAB19-HER2 interaction
01:13
27. Negative cooperative binding behaviour
01:15
28. Project II: Fcab-VEGF interaction: Generation of a two-faced Fcab
00:12
29. X-ray structure of CT6
00:32
30. X-ray structure of CT6
00:52
31. VEGF
01:05
32. Targeting VEGF
00:32
33. SEC-MALS
00:15
34. SEC-MALS
00:49
35. SEC-MALS
00:28
36. Generation of heterodimeric CT6
00:59
37. Generation of heterodimeric CT6
00:05
38. SEC-MALS
00:15
39. SEC-MALS
00:25
40. X-ray structure of the JanusCT6-VEGF complex
00:31
41. ITC measurement of JanusCT6-VEGF interaction
01:03
42. Summary & Conclusion
03:17
43. Summary & Conclusion
00:46
44. Thank You...
00:08
45. Thank You...
00:08
46. Question & Answer
00:39