Optimized variants of anti-VEGF antibodies
An antibody, FR-L1 technology, applied in the direction of antibodies, antibody medical components, antibody mimics/stents, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0556] Example 1: Deep-scanning mutagenesis of G6.31 identifies positions important for assembly of stably assembled Fabs
[0557] G6.31 is a high affinity anti-VEGF antibody (Fuh et al., J.Biol.Chem. 281(10):6625-6631 (2006); Liang et al., J.Biol.Chem.281(2): 951-961 (2006). Since the heavy and light chain variable domains of G6.31 are of common human germline IGHV3-23 and IGKV1-39 origin, respectively, they were considered to represent human antibodies.
[0558] To systematically assess the effect of single mutations in the VH and VL domains of G6.31, single sites of saturation were generated for each variable domain (VL positions 2-107 and VH positions 2-113, following the Kabat numbering scheme) Mutagenesis library. These libraries are referred to as VH and VL NNK walk libraries, respectively. The use of these libraries in deep mutation scanning experiments allowed the calculation of the enrichment ratio (ER) for each mutation in the two libraries during the selection pr...
Embodiment 2
[0572] Example 2: Deep scan mutagenesis screening of G6.31 identifies amino acid residue variants with increased stability and / or increased binding affinity for VEGF
[0573] In order to gain a comprehensive understanding of the impact of a single G6.31 substitution on antigen binding, the VH and VLNNK walk libraries described in Example 1 were panned against VEGF ( Figures 4A-4B ). The obtained enrichment ratio (ER) values showed a bimodal distribution ( Figures 5A-5B ). A subset of mutations had a strong negative effect on binding function, while the majority of mutations were neutral, and a small number of mutations had a strong positive effect on fitness. Mutations that have a negative effect on binding are largely located in HC-HVR due to the fact that these loops artificially provide most of the binding function of G6.31, or are located in conserved residues of the framework, as identified in anti-gD panning (see Example 1). Strongly enriched mutations were mainl...
Embodiment 3
[0593] Example 3: The conformational change of LC-F83 correlates with the Fab elbow conformation in the G6 structure
[0594] To understand how far a mutation is spatially distant from the antigen binding site can have such a strong effect on antigen binding and stability, the structural effect of the LC-F83A mutation was examined in more detail. The parental antibody G6 of G6.31 has previously been crystallized in VEGF-bound and VEGF-free forms (Fuh et al., supra). Compared to G6, G6.31 only carries four substitutions in HVR-L3. Therefore, the crystal structure of G6 was used as a model for G6.31. The crystal structure of the VEGF-free form of G6 contains 12 Fab molecules in an asymmetric unit. Based on the square shape (V-C interface) of the constant domains to the variable domains, the Fab structures can be clustered into two distinct groups ( Figure 6A ), which is the result of the different configurations of the Fab elbow region (elbow region), and can be quantified b...
PUM
Property | Measurement | Unit |
---|---|---|
T m | aaaaa | aaaaa |
Radius | aaaaa | aaaaa |
Radius | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com