摘要
目的：以重组人碱性成纤包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。半对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Ｗｅｓｔｅｒｎ ｂｌｏｔ法和免疫沉淀法对抗体特异性和ｌｇ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。
objective: Five hybridoma cell lines secreting anti-bFGF mAb have been developed from immunized Balb/c mice with recombinant human bFGF, by cell fusion. The variable region of K chain (Vk) genes was isolated by RT-PCR and sequencd. Methods:Two of these hybridoma cell lines were identified by using westem blot analysis. The recombinant vectors containing 4a2 Vk gene insert were identified by sequencing with Sanger's dideoxy-mediated chain termination method and computer-assisted comparative sequence analysis. Results: It showed that Vk gene consisted of 333bp encoding 111 amine acid resides. Comparing with EMBL and Kabat database, the Vk was in agree ment with the characterization of DNA sequence present in the mouse Ig Vk region. Conclusion:The success of cloning of the Vk gene of 4a2 McAb lay a good foundation for the consruction and expression of single chain variable fragment of 4a2 McAb.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2000年第7期353-355,共3页
Chinese Journal of Immunology
基金
国家"九五"科技重点攻关项目资助!96-C02-01-03
