title: Engineering the male-specificity of Fab against SDM antigen by chain shuffling.
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 κ 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.
authors.affiliation: College of Veterinary Medicine, Hunan Agricultural University, Changsha, Hunan Province, PR China.
authors.forename: Naidong
authors.initials: N
authors.lastname: Wang
authors.forename: Anwen
authors.initials: A
authors.lastname: Yuan
authors.forename: Zhibang
authors.initials: Z
authors.lastname: Deng
authors.forename: Qing
authors.initials: Q
authors.lastname: Yang
authors.forename: Jun
authors.initials: J
authors.lastname: Ma
authors.forename: Qinghui
authors.initials: Q
authors.lastname: Tan
authors.forename: Sizhe
authors.initials: S
authors.lastname: Zhang
authors.forename: Liqun
authors.initials: L
authors.lastname: Xue
authors.forename: Shuliang
authors.initials: S
authors.lastname: Cui
mesh_terms: D000609:Aminoacridines / Q000276:immunology*; D000818:Animals; D000918:Antibody Specificity / Q000235:genetics; D000941:Antigens / Q000276:immunology; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D005260:Female; D015321:Gene Rearrangement / Q000235:genetics; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D015202:Protein Engineering / Q000379:methods*; D011995:Recombination, Genetic / Q000502:physiology; D012727:Sex Characteristics
chemical_list: D000609:Aminoacridines; D000941:Antigens; D007140:Immunoglobulin Fab Fragments; C066906:SDM