A cell-cell interaction format for selection of high-affinity antibodies to membrane proteins.
Proc Natl Acad Sci U S A
; 116(30): 14971-14978, 2019 07 23.
Article
em En
| MEDLINE
| ID: mdl-31285332
ABSTRACT
Generating and improving antibodies and peptides that bind specifically to membrane protein targets such as ion channels and G protein-coupled receptors (GPCRs) can be challenging using established selection methods. Current strategies are often limited by difficulties in the presentation of the antigen or the efficiency of the selection process. Here, we report a method for obtaining antibodies specific for whole cell membrane-associated antigens which combines a cell-cell interaction format based on yeast display technology with fluorescence-activated cell sorting of dual fluorescent complexes. Using this method, we were able to direct the affinity maturation of an antagonist antibody specific for the proton-gated ion channel ASIC1a and showed that both the affinity and potency were improved. We were also able to use this method to do kinetic selections to generate clones with better dissociation profiles. In addition, this method was employed successfully to handle the difficult problem of selecting antibodies specific to a GPCR target, the mu-opioid receptor.
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Base de dados:
MEDLINE
Assunto principal:
Receptores Acoplados a Proteínas G
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Descoberta de Drogas
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Citometria de Fluxo
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Canais Iônicos
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Anticorpos
Limite:
Animals
Idioma:
En
Ano de publicação:
2019
Tipo de documento:
Article