鸡源性免疫单链抗体库的构建与鉴定

Construction and identification of an immune single chain antibody library derived from a chicken

目的:建立鸡源性免疫单链抗体库的构建方法。方法:根据鸡抗体基因结构特点,借助噬菌体展示技术原理,以免疫变形链球菌全菌抗原后的鸡脾脏B细胞为抗体基因来源,构建鸡源性免疫单链抗体库;采用固相淘选法筛选抗变形链球菌的噬菌体单链抗体(scFv),以此鉴定所构建的免疫单链抗体库。结果:从鸡脾脏总RNA中扩增出了鸡免疫球蛋白的轻、重链可变区的全部基因;得到了一个库容量为1·6×107的鸡免疫单链抗体库;筛选到了抗变形链球菌的噬菌体scFv,并测出了其DNA序列。结论:结合噬菌体抗体库技术原理与鸡抗体基因结构的特点,可以成功地构建鸡源性免疫单链抗体库。

AIM： To develop a method for constructing an immune single chain antibody library derived from a chicken. METHODS： According to the principle of phage display and the character of chicken antibody genes, an im- mune single chain antibody library was constructed by using the spleen B cells which were extracted from a chicken im- munized with streptococcus mutans as antibody gene sources. According to the principle of immobilized panning the specific chicken single chain phage antibody was select- ed from the single chain antibody library to identify the immune antibody library. RESULTS： The repertoire of the chicken heavy-chain and light-chain variable regions was obtained from the total intact RNA of chicken spleen. The phage antibody library was generated and its size was determined to be 1.6×10^7 A specific chicken single chain phage antibody against streptococcus mutans was selected and its DNA was sequenced. CONCLUSION： An immune single chain antibody library derived from a chicken can be constructed successfully based on the principle of phage antibody library and the character of chicken antibody genes.