Optimization and Modification of Anti-rhTNF-α Single Chain Variable Fragment Antibody: Effective In Vitro Affinity Maturation and Functional Expression of Chimeric Fab

Xiaoniu, M, et al., 67(5), 437-444, Biomed Pharmacother, 2013

The CDR-H3 domain of a well-characterized anti-rhTNF-α scFv fragment was mutated by degenerate PCR and the mutant library screened for higher affinity binders by phage display. An improved clone, scFv-G10, was identified and used along with constant regions CH1 and CL of human IgG1 to construct a novel vector for expression of a functional chimeric Fab. Biotinylated rhTNF-α was immobilized onto Streptavidin biosensors and binding affinity of scFv-G10 as compared to the original parent scFv clone was determined using the Octet RED system.

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