摘要
鼠单克隆抗体AA98是我室研制的一株新型抗肿瘤血管抗体 ,体内实验证明它可以抑制血管生成 ,抑制肿瘤生长。从分泌抗体AA98的杂交瘤细胞中提取总RNA ,采用逆转录 PCR(RT PCR)分别扩增重链Fd及轻链κ,并进行序列测定。将Fd和κ链依次与噬菌粒 pComb3H连接 ,构建pComb3H AA98Fab表达载体。在大肠杆菌中表达了AA98Fab。免疫印迹表明该Fab片段识别分子量为 10 0kD的蛋白 ,具有原抗体AA98的抗原特异性。AA98Fab是研究抗体AA98作用机理的工具 ,也为制备重组免疫毒素 。
A tumor blood vessel specific antibody fragment Fab was constructed and expressed in E.coli. Our lab had generated a mouse monoclonal antibody,referred to as AA98. Targeting tumor blood vessel,AA98 potently inhibited tumor growth and metastasis in vivo . Total RNA was extracted from hybridoma secreting AA98 and cDNA was then synthesized. Using PCR,heavy chain and light chain of AA98 were amplified and ligated into phagemid pComb3H to construct recombinant expression vector pComb3H Fab. Soluble Fab was then expressed in E.coli XL1 Blue. Immuno\|blotting showed that the expressed Fab could recognize a 100kD band, maintaining the same specificity of its parent antibody AA98. This recombinant FAb fragment is an important reagent to investigate the mechanism of AA98 inhibiting tumor angiogenesis and to develop recombinant AA98 conjugated immuno\|toxin for tumor blood vessel targeted therapy.
出处
《生物工程学报》
CAS
CSCD
北大核心
2000年第6期695-698,共4页
Chinese Journal of Biotechnology
基金
国家高技术研究发展与计划项目!( 10 2 0 9 0 2 0 4)
