Antibody Batch Cloning.
Methods Mol Biol
; 2702: 411-417, 2023.
Article
en En
| MEDLINE
| ID: mdl-37679632
ABSTRACT
The antigen-binding ability of each antibody clone selected by phage display is usually initially ranked by a screening ELISA using monovalent scFv antibody fragments. Further characterization often requires bivalent antibody molecules such as IgG or scFv-Fc fusions. To produce these, the V region encoding genes of selected hits have to be cloned into a mammalian expression vector and analyzed as a bivalent molecule, requiring a laborious cloning procedure. We established a high-throughput procedure allowing rapid screening of candidates in bivalent formats. This protocol allows for the parallelized cloning of all selected antibody fragments into a mammalian expression vector in the 96-well plate format. The bivalent antibody molecules can then be produced and purified in 96-well plates for further analysis in microtiter plate assays.
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1
Banco de datos:
MEDLINE
Asunto principal:
Fragmentos de Inmunoglobulinas
/
Anticuerpos
Límite:
Animals
Idioma:
En
Año:
2023
Tipo del documento:
Article