Synthetic polynucleotides libraries
A technology for synthesizing polynucleotides and polynucleotides, applied in the field of synthetic polynucleotide libraries, can solve the problems of underrepresentation, lack, becoming immunogenic, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment approach
[0261] While the methods described herein demonstrate the production of theoretical segment pools of H3-JH and DH segments using a limited number of allelic variants, those skilled in the art will recognize that the methods taught herein may be applicable to any IGHJ and IGHD gene, including any Other allelic variants and all non-human IGHJ and IGHD genes. Alternatively or additionally, the methods described herein may be applied to any reference set of CDRH3 sequences, eg to extract additional TN1 and / or N2 segments. Alternatively or additionally, those skilled in the art will recognize that each embodiment described herein may be in the form of a polynucleotide or polypeptide) within a vector, virus, or microorganism (eg, yeast or bacteria). Furthermore, since the invention is fully recited in relation to synthetic libraries, certain embodiments of the invention and uses thereof relate to any of the embodiments described above, in computer readable format.
[0262] Librarie...
Embodiment 1
[0277] Example 1. Light chain libraries with framework and / or CDRL1 and / or CDRL2 variability
[0278] Although the diversity of antibody sequences is concentrated in the CDRs, certain residues within the framework regions can also affect antigen recognition and / or modulate affinity (Queen et al., Proc. Natl. Acad. Sci. USA, 1989, 86: 10029; Carter et al., Proc. Natl. Acad. Sci. USA, 1992, 89:4285, all incorporated by reference in their entirety). These residues have been cataloged and used to make framework substitutions to increase the affinity of antibodies, for example, during antibody humanization (see for example "Vernier" residues, Foote and Winter, J. Mol. Biol., 1992, 224:487 , incorporated by reference in its entirety). In the heavy chain, Vernier residues include Kabat-numbered residues 2, 27-30, 47-49, 67, 69, 71, 73, 78, 93-94, and 103. In the light chain, Vemier residues include Kabat residues 2, 4, 35-36, 46-49, 64, 66, 68-69, 71, and 98. The number of Vernier...
Embodiment 2
[0312] Example 2. A light chain library with enhanced diversity in CDRL3
[0313] Various methods of making light chain libraries are known in the art (eg, see US Publication Nos. 2009 / 0181855, 2010 / 0056386, and WO / 2009 / 036379). Analysis of clinically validated antibody sequences revealed rearrangements from germline-like VL-JL (where "L" can be a kappa or lambda germline sequence), in the presence of somatic mutations ( figure 2 ) before, the deviations of these sequences are very small. Here, a germline-like rearrangement is one in which either the V or J portion differs from the respective germline gene, and for the purposes of this specific example, wherein the length of CDRL3 is restricted to 8, 9 or 10 amino acids (see US Publication Nos. 2009 / 0181855, 2010 / 0056386, and WO / 2009 / 036379). However, for the IGHJK1 gene, the WT (Trp-Thr) and RT (Arg-Thr) sequences (the first 2 N-terminal residues) were considered "germline-like" and a complete L3 rearrangement contained th...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More