Systems and methods to classify antibodies
A technology for antigen-binding molecules and antigens, applied in the field of systems and methods for classifying antibodies, capable of solving problems such as reducing antigen binding
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[0106] This example describes an exemplary application of the systems and methods described herein to the CDRH3 of the trastuzumab (Herceptin) antibody and to classify the antibody binding to the corresponding target HER2 antigen.
[0107] A. Results
[0108] 1) Deep mutation scanning to determine the antigen-specific sequence landscape (landscape) and guide rational antibody library design
[0109] Since the amino acid sequence of CDRH3 of an antibody is a key determinant of antigen specificity, deep mutation scanning (DMS) was performed on this region to resolve the specificity-determining residues. First, a hybridoma cell line expressing a trastuzumab variant unable to bind the HER2 antigen (mutated CDRH3 sequence) was used ( FIG. 9 ). Libraries were generated by CRISPR-Cas9-mediated homology-directed mutagenesis (HDM) (Mason et al. (2018) Nucleic Acids Research 46(14):7436–49) utilizing the CDRH3 The Cas9-targeted gRNA and the pool of homologous templates in the form of ...
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