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目的：以重组人碱性成纤维细胞生长因子为抗原免疫小鼠，建立多株稳定分泌bFGF单克隆抗体（McAb）的杂交瘤细胞。并对bFGF单克隆抗体（McAb）可变区基因进行扩增及序列分析。方法：采用Westernblot法和免疫沉淀法对抗体特异性和IG类型进行鉴定，同时，应用分子生物学技术，对Vk基因的DNA片段进行克隆，并对4a2Vk基因进行序列测定。结果：bFGF单克隆抗体可特异性结合人重组bFGF抗原，且均为IgM。由杂交瘤4a2、3d3细胞株扩增Vk基因是由330个碱基组成，编码110个氨基酸残基。通过国际联机检索进行EMBL和Kaba基因库扫描发现，4a2VkDNA仅与Ig同源，符合小鼠Ig的Vk基因特征；同源性为90％~98％，应归属于Ig的Vk基因。结论：杂交瘤4a2、3d3细胞株可稳定分泌bFGF单克隆抗体，并获得其轻链可变区基因，为进一步构建和表达单链Fv（ScFv）抗体打下良好的基础。
OBJECTIVE: To immunize mice with recombinant human basic fibroblast growth factor (FGF) as antigen and establish multiple hybridoma cells stably secreting bFGF monoclonal antibody (McAb). The bFGF monoclonal antibody (McAb) variable region gene was amplified and sequenced. Methods: Antibody specificity and IG type were identified by Western blot and immunoprecipitation. At the same time, the DNA fragment of Vk gene was cloned by molecular biology techniques and the sequence of 4a2Vk gene was sequenced. RESULTS: The bFGF monoclonal antibody specifically binds to human recombinant bFGF antigen and is IgM. Amplification by Hybridoma 4a2 and 3d3 Cell Lines The Vk gene consists of 330 bases and encodes 110 amino acid residues. The international online search of EMBL and Kaba gene bank found that 4a2VkDNA was only homologous with Ig, which was in accordance with the characteristics of mouse Ig Vk gene. The homology was 90% ~ 98% and should belong to Vk gene of Ig. CONCLUSION: The hybridoma 4a2 and 3d3 cell lines can stably secrete monoclonal antibody against bFGF and obtain the light chain variable region gene, which laid a good foundation for further construction and expression of single chain Fv (ScFv) antibody.