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目的本研究以原核表达单抗原表位的人心肌红蛋白(MB)肽段,制备不同表位的多克隆抗体。方法利用软件分析人心肌红蛋白的抗原表位,获得4个肽段(片段1、2、3、4);PCR扩增其基因序列,连接到表达载体p ET-GST,转化至E.coli BL21进行诱导表达。结果 GST-Sepharose 4B亲和层析柱进行MB-GST融合蛋白纯化,获得Mr28 000左右的融合蛋白,质量浓度为15 mg/L。免疫新西兰大白兔获得高效价的多克隆抗体。结论 4种多肽抗体均具有较好的特异性。
OBJECTIVE In this study, human monoclonal myelopeptide (MB) fragment was expressed in prokaryotic cells to prepare polyclonal antibodies with different epitopes. Methods Four peptides (fragments 1, 2, 3 and 4) were obtained by software analysis of human cardiac myoglobin. The gene sequences were amplified by PCR and ligated into expression vector p ET-GST and transformed into E. coli BL21 induced expression. Results The GST-Sepharose 4B affinity column was purified by MB-GST fusion protein, and the fusion protein Mr28 000 was obtained with a mass concentration of 15 mg / L. Immunization of New Zealand white rabbits to obtain high titer of polyclonal antibodies. Conclusion The four kinds of peptide antibodies have good specificity.