从大容量噬菌体抗体库中筛选人源性抗角蛋白scfv-首都医科大学学报.pdfVIP

2007 2 F eb. 2007 2 8 1 Journal of C apitalM ed ical U niversity V o l. 28 N o. 1 从大容量噬菌体抗体库中筛选人源性抗角蛋白 scFv 1 2 2 1 1* 张国民 乔媛媛 陈宇萍 吕 喆 安云庆 1. ; 2. ) ! scFv) , , , 4 ∀ #, , EL ISA , DNA 4 , 9 , , DNA , ! ; ; ! R 392. 12 Cloning of Antikeratin ScFv F rom a H um an Large Phage Antibody L ibrary 1 2 2 1 1* Zhang Guom in , Q iao Yuanyuan , Chen Yuping , LZhe , A n Y unq ing 1. D ep artment of M icrobiology and Immunology, Cap italM edical University; 2. CentralLab of Navy GeneralHosp ital) AB STRA CT! O b ective T o c lone hum an sing le cha in an tibod ies scFv ) ag ainst ke ratin from a large phage an tibody library and analyze the spec ific ity of antigen b inding activ ity and the hom o logy of gene sequences. M e thods A hum an la rge phage antibody library w as panned for four tmi es. The spec if ic ity of the antigen b inding activ ity of the se lected c lones w as identif ied by EL ISA. Positive c lones w ere ana lyzed by DNA f ingerpr int and sequenc ing. R esu lts A fter 4 rounds of ∀ adhesione lutionam plif ica tion# panning, 9 c lones expressing anti keratin scFv w ere ob tained. DNA f ingerprint and sequence ana lys is show ed tha t they had different antibody V reg ion encod ing g enes. Con clusion H um an an ti kera tin scFv is obta ined from a large phage antibody library, w h ich w ill bene fit to the app lication of antikeratin antibody to clin ica l research. KEY W ORDS! phage antibody libra ries; keratin; scFv scFv, , , 1 AK autoA b), [ 1] 1. 1 mA b),