对一株基因工程人抗角蛋白抗体的免疫学鉴定

Immunologic Characterization of a Genetically Engineered Human Antibody Against Keratin

目的对1株基因工程人抗角蛋白抗体识别抗原的特性进行鉴定。方法利用从噬菌体抗体库中筛选到的特异表达抗角蛋白的Fab片段的质粒转化大肠杆菌,IPTG诱导表达出Fab片段,用ELISA鉴定抗原结合活性和特异性,用蛋白免疫印迹和竞争抑制性ELISA检测抗体所识别的角蛋白组分。结果该株基因工程人抗角蛋白抗体的结合活性和特异性良好,所识别的抗原为相对分子质量46000角蛋白,为特异性人抗角蛋白K17抗体。结论该抗体具有优良的免疫学特性,有必要对其生物学活性和临床应用前景做进一步探讨。

Objective To immunologically characterize the binding activity of a genetic engineering human antibody against keratin. Methods The specific anti-keratin Fab was screened from a phage antibody library. After Fab monoclonal antibody was induced by isopropylthio-β-D-galactoside (IPTG), the binding activity with antigens and antigenic specificity of the antibody were verified by ELISA,Western blot, and competitive inhibition ELISA. Results The antibody possessed good antigenic specificity as well as excellent binding activities with antigens, and could recognize 46 000 keratin specifically. It was identified as anti-keratin 17 antibody. Conclusions The immunologic characteristics of the genetic engineering antibody are very good. It is necessary to further study the biological activities and clinical application of it.