1. Engineering the male-specificity of Fab against SDM antigen by chain shuffling.
- Author
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Wang N, Yuan A, Deng Z, Yang Q, Ma J, Tan Q, Zhang S, Xue L, and Cui S
- Subjects
- Animals, Antigens immunology, Cells, Cultured, Female, Gene Rearrangement genetics, Immunoglobulin Fab Fragments biosynthesis, Immunoglobulin Fab Fragments metabolism, Male, Mice, Mice, Inbred C57BL, Recombination, Genetic physiology, Sex Characteristics, Aminoacridines immunology, Antibody Specificity genetics, Cloning, Molecular methods, Immunoglobulin Fab Fragments genetics, Protein Engineering methods
- Abstract
High-titer serologically detected male (SDM) antibody fragments are essential for specific binding to the SDM antigen and promoting its application. The A8 clone previously obtained from an original phage antibody library was further affinity-matured by light- and high-chain shuffling respectively, to generate the end product B9 clone. The binding capacity of B9 phage Fabs to male splenocytes doubled the value of its parental A8 clone (determined using ELISA). Based on immunofluorescent staining, B9-Fabs mainly bound to the surface antigen of male splenocytes and recognized testicular cells. The resulting B9-Fabs detected a single protein (approximately 40 kDa determined using Western blot analysis of male splenocytes and testis); its high SDM antigen binding ability might have been because of mutation sites and varied lengths of the amino acid sequences in the complementarity determining regions-3 of the Îș and Fd chains. The new recombinant clones of Fab that were phage-enhanced using chain shuffling were candidate molecules for investigating molecular mechanisms of SDM antigens specific binding and applications., (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Published
- 2013
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