PCR条件及程序改变对抗体库多样性的影响  

Effect of Different PCR Procedures and Conditions to the Diversity of Humanized Phage Display Antibody Library.

作　　者：田学军[1,2] 寿成超[1,2] 孟麟[1,2] 董志伟[1,2] 

机构地区：[1]北京医科大学临床肿瘤学院 [2]中国医学科学院肿瘤研究所

出　　处：《生物化学与生物物理进展》1999年第2期172-176,共5页Progress In Biochemistry and Biophysics

基　　金：国家"863"高技术项目资助

摘　　要：用家族特异性免疫球蛋白可变区基因引物两两组合分别对轻、重链进行ＲＴＰＣＲ扩增．在不同退火温度下得到了全部轻链及大部分重链（１３／１６）可变区基因．当先用免疫球蛋白信号肽序列５′端引物与未扩出基因３′端引物组合进行一次ＰＣＲ，再以该产物为模板进行二次ＰＣＲ，则获得了未扩出三条重链的ＰＣＲ产物．这表明通过ＰＣＲ反应条件的改变及程序调整，可增加所获得可变区基因的种类及数量。Human immunoglobulin heavy chain and light chain genes were separately amplied by PCR with cDNA as templates from human peripheral lymphocytes ,using four 5′ primers and four 3′ primers corresponding to heavy chain, eight 5′ primers and two 3′ primers corresponding to light chain. At different annealing temperature, using the primers described above, all the light chain PCR products and 80% heavy chain PCR products were yielded. When signal sequences of immunoglobulin were used as 5′ primers, all remain heavy chain products were obtained by this half nested PCR. This results indicate that more immunoglobulin variable region genes can be obtained by PCR through changed of procedures and reaction conditions, which can increased the diversity of antibody library.

关 键 词：退火温度 抗体库 多样性 半套式PCR 

分 类 号：R392.12[医药卫生—免疫学] R78[医药卫生—基础医学]