Augmented Binary Substitution: Single-pass CDR germ-lining and stabilization of therapeutic antibodies
| Autor: | Orla Cunningham, Janet E. Paulsen, Barry McDonnell, Lioudmila Tchistiakova, Deirdre Ní Shúilleabháin, Joanne Grant, Sue Townsend, Alfredo Darmanin-Sheehan, Niall J. Foy, Conor Fields, Jason Wade, Brian J. Fennell, James R. Apgar, Amy King, Matthew Allister Lambert, William J.J. Finlay, Edward Franklin, Timothy P. Hickling |
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| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Models
Molecular Molecular Sequence Data Immunoglobulin Variable Region Epitopes T-Lymphocyte Sequence alignment Enzyme-Linked Immunosorbent Assay tau Proteins Complementarity determining region Computational biology Biology Immunoglobulin G Antibody Specificity Peptide Library Sequence Analysis Protein Animals Humans Computer Simulation Amino Acid Sequence Peptide library Multidisciplinary Protein Stability Immunogenicity Antibodies Monoclonal Biological Sciences Complementarity Determining Regions Clone Cells Protein Structure Tertiary Rats Germ Cells Immunology Mutation biology.protein Immunoglobulin heavy chain Paratope Immunoglobulin Light Chains Antibody Immunoglobulin Heavy Chains Sequence Alignment |
| Popis: | Although humanized antibodies have been highly successful in the clinic, all current humanization techniques have potential limitations, such as: reliance on rodent hosts, immunogenicity due to high non-germ-line amino acid content, v-domain destabilization, expression and formulation issues. This study presents a technology that generates stable, soluble, ultrahumanized antibodies via single-step complementarity-determining region (CDR) germ-lining. For three antibodies from three separate key immune host species, binary substitution CDR cassettes were inserted into preferred human frameworks to form libraries in which only the parental or human germ-line destination residue was encoded at each position. The CDR-H3 in each case was also augmented with 1 ± 1 random substitution per clone. Each library was then screened for clones with restored antigen binding capacity. Lead ultrahumanized clones demonstrated high stability, with affinity and specificity equivalent to, or better than, the parental IgG. Critically, this was mainly achieved on germ-line frameworks by simultaneously subtracting up to 19 redundant non-germ-line residues in the CDRs. This process significantly lowered non-germ-line sequence content, minimized immunogenicity risk in the final molecules and provided a heat map for the essential non-germ-line CDR residue content of each antibody. The ABS technology therefore fully optimizes the clinical potential of antibodies from rodents and alternative immune hosts, rendering them indistinguishable from fully human in a simple, single-pass process. |
| Databáze: | OpenAIRE |
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