manual Ph.D. Phage Display Libraries E8100, E8101, E8102 ...
XOMA NON-Confidential Phage libraries 2017 Non... · 2018-08-22 · XOMA’s Phage Display...
Transcript of XOMA NON-Confidential Phage libraries 2017 Non... · 2018-08-22 · XOMA’s Phage Display...
Phage Display Libraries
XOMAAntibody Platform Technologies ExpandTarget Opportunities while Generating Antibodies with Novel Properties
▪ The XOMA antibody platform is built as a combination of novel technologies and unique know-how
▪ XOMA’s access to major commercial phage libraries provides knowledge and experience for creating top-performing custom libraries
▪ Novel expression-enhancing advancements improve the functional activity of all phage libraries
▪ Optimized panning techniques developed at XOMA maximize specific hits to cell-surface targets, including GPCRs, and help ID antibodies with unique modulatory function
▪ Novel yeast and mammalian display platforms accelerate Ab discovery by allowing rapid IgG reformatting and FACS-based antibody screening
The XOMA antibody platform accelerates antibody discovery and is making difficult cell‐surface targets and mechanisms of action accessible
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XOMA’s Phage Display Libraries have SeveralXOMA Phage Display Technology
• XOMA089 TGFb: Ph1• Licensed to Novartis
• XOMA358 insulin receptor: Ph2• Congenital
Hyperinsulinism• Post-Gastric Bypass
Hypoglycemia
Clinical Validation
XOMA’s Phage Display Libraries have SeveralCompetitive Advantages
XOMA has some of the most diverse phage display antibody libraries• Picomolar affinity antibodies routinely selected with limited screening• Antibody fragments with different target specificities and diverse epitope coverage can
be isolated from a single panning campaign
Competitive Advantages of XOMA’s Technology
Vector Design Library Structure Freedom to Operate2 31
• Three tags (V5‐6His‐c‐myc)
• Amber codon
• Gene 3 stump
• Very large (>1011)
• Fully human
• Natural repertoire
• Choice of fragment format
• High percentage of open reading frames
• No gatekeeping
• Unique IP package
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XOMA031 Fab Library Characteristics
▪ 30 healthy donors▪ 14 PBMC, 15 bone marrow, 1 lymph node
▪ Total library size: 3.4 x 1011
▪ Kappa sub‐library: 1.8 x 1011
▪ Lambda sub‐library: 1.6 x 1011
▪ Full human repertoire▪ All VH, VL, and VK families
▪ Expression▪ 69% of Kappa clones express full length fragments
▪ 81% of Lambda clones express full length fragments
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Human XOMA031
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2020Library Validation Summary for 3 Panning Rounds
XOMA031 Fab LibraryTarget Panning
MethodPlates
ScreenedHit Rate Total Unique Unique VH Unique VL
Gastrin Soluble 5 38% 21 16 17
FITC-BSA Immobilized 6 41% 108 103 102
Transferrin Soluble 1 55% 35 27 35
TIE-2 Soluble 4 51% 66 40 37
XOMA040 scFv Library Characteristics
▪ 33 healthy donors▪ 13 PBMC, 5 bone marrow, 13 spleen and 2 thymus
▪ Total library size: 3.8 x 1011
▪ Kappa sub‐library: 1.38 x 1011
▪ Lambda sub‐library: 2.44 x 1011
▪ Full human repertoire▪ All VH, VL, and VK families
▪ Expression▪ 71% of Kappa clones express full length fragments
▪ 71% of Lambda clones express full length fragments
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Human XOMA040
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15kDa soluble protein
Panning and Screening Funnel
3 rounds of soluble panning with a biotinylated 15kDa protein
Primary Screen of 4 96‐well plates as PPE 1) ELISA – 176/372 (47%)2) FACS (Target bound to cell surface receptor) – 165/372 (44%) 3) SPR – 197/372 (53%)
FACS secondary screen of 325 clones (hit in any of the primary screens)1) Target bound to monomeric receptor 12) Target bound to dimeric receptor 1/23) Target bound to monomeric receptor 3
Sequenced 214 clones (hit in secondary screens 1 and/or 2 but not 3)
82 mAbs reformatted
78 mAbs expressed as IgG2
72 mAbs retained binding as IgG2
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▪ 82 unique clones
▪ 66 unique VH
▪ 78 unique VL
▪ 41 Kappa LC
▪ 37 Lambda LC
Family distribution of selected clones
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▪ 72 mAbs with affinities between 1µM and 10pM
▪ 34 mAbs < 10nM
▪ 10 mAbs < 1nM
High Affinity clones discovered from XOMA040 without affinity maturation
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Library Validation Summary for 3 Panning Rounds
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XOMA031 Fab Library
XOMA040 scFv Library
Target Panning Method
Plates Screened
Hit Rate Total Unique Unique VH Unique VL
Gastrin Soluble 5 38% 21 16 17
FITC‐BSA Immobilized 6 41% 108 103 102
Transferrin Soluble 1 55% 35 27 35
TIE‐2 Soluble 4 51% 66 40 37
Target Panning Method
Plates Screened
Hit Rate Total Unique Unique VH Unique VL
ANG‐2 Soluble 6 96% 59 41 50
FITC‐BSA Immobilized 2 96% 134 105 107
XOMA Library Summary
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▪ XOMA040 and XOMA031 utilize all natural frameworks for increased probability of isolating a diverse set of functional antibodies.
▪ All XOMA libraries consistently deliver diverse sets of high affinity antibodies