文章摘要
毕立清,刘 莉,程蕴琳,施有为,仇镇宁,姜苏蓉.抗人胰岛素样生长因子-1单克隆抗体的制备与鉴定[J].南京医科大学学报,2008,28(6):712~716741
抗人胰岛素样生长因子-1单克隆抗体的制备与鉴定
Preparation and identification of monoclonal antibody against human insulin-like growth factor-1
投稿时间:2007-09-29  
DOI:10.7655
中文关键词: 人胰岛素样生长因子-1  单克隆抗体  重组蛋白
英文关键词: human insulin-like growth factor-1  monoclonal antibody  recombinant protein
基金项目:江苏省卫生厅"135"重点医学工程基金(135-14);江苏省卫生厅科技发展基金(H200511);江苏省卫生厅"十一五兴卫工程/老年医学重点学科"专项基金
作者单位
毕立清 南京医科大学第一附属医院老年医学科,江苏 南京 210029 
刘 莉 南京医科大学第一附属医院老年医学科,江苏 南京 210029 
程蕴琳 南京医科大学第一附属医院老年医学科,江苏 南京 210029 
施有为 南京医科大学第一附属医院老年医学科,江苏 南京 210029 
仇镇宁 南京医科大学卫生部抗体技术重点实验室,江苏 南京 210029 
姜苏蓉 南京医科大学第一附属医院老年医学科,江苏 南京 210029 
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中文摘要:
      目的:制备稳定分泌抗人胰岛素样生长因子-1(hIGF-1)单克隆抗体(mAb) 的杂交瘤细胞系,并对其分泌的mAb进行鉴定?方法:用纯化后的hIGF-1重组蛋白免疫Balb/c小鼠,利用杂交瘤细胞融合技术制备抗hIGF-1的mAb,用Western blot法鉴定mAb的特异性,用间接ELISA法检测mAb腹水效价及mAb的亲和力,杂交瘤细胞染色体分析,鉴定Ig亚类?结果:获得2株能稳定分泌抗hIGF-1的mAb杂交瘤细胞系,Western blot分析显示,该mAb能与具有生物学活性的成熟IGF-1蛋白结合?腹水mAb效价可达6 × 104?mAb相对亲和力为2.1×109/mol~7.8×109/mol?2株单抗属IgM亚类?染色体分析符合杂交瘤细胞特征?结论:成功制备出抗hIGF-1的2株mAb杂交瘤细胞,为进一步用于hIGF-1的临床检测和实验研究创造了条件?
英文摘要:
      Objective:To establish the specific hybridoma cell lines that stably secreted monoclonal antibody(mAb) against human insulin-like growth factor-1 and identificate its properties. Methods:Balb/c mice were immunized with purified recombinant human IGF-1 protein. Splenocytes of the immunized mice were collected and fused with the mouse myeloma cell line Sp2/0 cells. The hybridoma cells that secreted human IGF-1 mAb were subcloned with limited dilution. The specificity of mAb was detected by Western blot. Indirect enzyme-linked immunosorbent assay(ELISA) was used to evaluate the affinity and titers of mAbs. The immunoglobulin (Ig) subtype of mAb was determined by test paper bar. Results:Two stably secreting anti-hIGF-1 mAb cell lines were obtained. Western blot analysis showed the mAb had specific combination with human IGF-1 which possessed biological activities. The titers of both mAb in ascites were 6 × 104 and their relative affinities were between 2.1 × 109/mol~7.8 × 109/mol. Two mAb cell lines can secret IgM subtype antibodies. Conclusion:Two hybridoma cell lines that secreted anti-hIGF-1 mAb were obtained,which will be useful for clinical application and fundamental basic research in hIGF-1.
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