抗猪流感病毒HA蛋白单克隆抗体重链基因的克隆及序列分析

Cloning and sequencing of the variable region fragments encoding the monoclonal antibody against the HA protein of swine influenza virus

为获得抗猪流感病毒HA蛋白单克隆抗体(MAb)重链可变区基因,提取MAb杂交瘤细胞8C4总RNA,通过RT-PCR扩增其重链可变区基因。其DNA序列与GenBank数据库和IMGT/V-QUEST软件比对,序列包含CDR1、CDR2、CDR33个高变区,CDR1含GYTFTSYY 8个氨基酸、CDR2含IYPGDGST 8个氨基酸、CDR3含ARVMIAMDY 9个氨基酸。第22位和96位为骨架区的2个半胱氨酸,形成链内特征性二硫键。该序列符合鼠Ig重链基本框架结构,框架区内无终止密码子,为重排产生的序列。本实验获得的重链序列为研制基因工程抗体奠定了物质基础。

The gene encoding heavy chain variable region of a monoclonal antibody （MAb） against the HA protein of swine influenza virus was amplified by RT-PCR from total RNA of the hybridoma cell secreting MAb 8C4. Sequence analysis showed that the VH gene contains 375 nucleotides, encoding 125 amino acids. There are 8 amino acids GYTFTSYY in CDRI, 8 amino acids IYPGDGST in CDR2, and 9 amino acids ARVMIAMDY in CDR3. The 22 and 96 cysteine formed typical disulfide bond in the VH gene. These data provided the foundation for the development of gene engineering antibodies.