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轮换淘选法筛选凝血酶特异的噬菌体抗体
引用本文:洪龙,杨柏成,朱圣庚. 轮换淘选法筛选凝血酶特异的噬菌体抗体[J]. 微生物学通报, 2005, 32(3): 73-76
作者姓名:洪龙  杨柏成  朱圣庚
作者单位:北京大学生命科学学院,北京,100871
基金项目:国家自然科学基金资助重点项目(No.20333010)~~
摘    要:以抗原为选择剂,通过淘选可以从噬菌体抗体库中获得特异的单链抗体克隆。采用液相一固相轮换淘选的方法,从鼠源噬菌体抗体库中淘选出与凝血酶特异结合的单链抗体。首先,用光敏生物素将凝血酶生物素化,然后用链亲和素磁珠法淘选与凝血酶特异结合的重组噬菌体。噬菌体扩增后使用酶标板进行第2轮淘选,以除去上一轮中非特异结合的重组噬菌体。经4轮轮换淘选,最后从23个单克隆噬菌体抗体中分离出4个凝血酶特异的噬菌体抗体。

关 键 词:轮换淘选法,噬菌体抗体库,凝血酶特异的单链抗体
文章编号:0253-2654(2005)03-0073-04
修稿时间:2004-09-06

The Selection of Thrombin-specific scAbs by Alternative Panning Strategy
Hong Long,YANG Bai-Cheng,ZHU Sheng-Geng. The Selection of Thrombin-specific scAbs by Alternative Panning Strategy[J]. Microbiology China, 2005, 32(3): 73-76
Authors:Hong Long  YANG Bai-Cheng  ZHU Sheng-Geng
Affiliation:Hong Long YANG Bai-Cheng ZHU Sheng-Geng~
Abstract:Specific scAbs could be obtained through biopanning from phage antibody libraries by use of antigens as target molecules. scAbs specific to thrombin were separated from mouse antibody library by the panning strategy of alternating liquid-solid phase in this paper. Thrombin was biotinylated by photobiotin at first, then avidin-coated magnetic beads were utilized to isolate specific scAbs. The eluted phages were amplified and subject to the second round panning in microtiter plate to remove the unspecific reombinant phages. 4 specific scAbs were separated from 23 phage clones after four rounds of alternative panning.
Keywords:Alternative panning strategy   Phage antibody library   Thrombin-specific scAb  
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