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目的:获得抗人视网膜母细胞瘤单克隆抗体轻链可变区基因。方法:从分泌抗人视网膜母细胞瘤单克隆抗体的杂交瘤细胞中提取总RNA,逆转录形成cDNA,用扩增小鼠K轻链可变区基因的引物,通过RT-PCR方法,扩增基因,1.5％琼脂糖凝胶电泳鉴定。结果:用轻链引物扩增获得340bp的基因片段。结论:从三株杂交瘤细胞中均获得高纯度的总RNA;用RT-PCR方法从一株杂交瘤细胞成功克隆了抗人视网膜母细胞瘤单克隆抗体轻链可变区基因,为基因工程抗体研究奠定了良好基础。眼科学报2001;17:194～197。
OBJECTIVE: To obtain the light chain variable region gene of monoclonal antibody against human retinoblastoma. METHODS: Total RNA was extracted from hybridoma cells secreting anti-human retinoblastoma monoclonal antibody, reverse transcribed to form cDNA, amplified with primers for the variable region gene of mouse K light chain by RT-PCR Gene, 1.5% agarose gel electrophoresis identification. Results: The 340bp gene fragment was amplified by light chain primer. Conclusion: The high purity of total RNA was obtained from all the three hybridoma cells. The variable region gene of monoclonal antibody against human retinoblastoma was cloned successfully from a hybridoma cell by RT-PCR. Antibody research has laid a good foundation. Journal of Ophthalmology 2001; 17: 194 ~ 197.