OLFM1蛋白的生物信息学分析及其多克隆抗体制备与鉴定

Bioinformatics analysis of OLFM1 epitope and preparation of polyclonal antibody

目的:利用生物信息学分析OLFM1蛋白,并制备其多克隆抗体。方法:根据生物信息学分析预测OLFM1蛋白的二级结构、亲水性、抗原性等理化特性,以这些分析预测为依据,经过同源性检索后设计出一段由20个氨基酸组成的多肽,免疫家兔,获得的多克隆抗体经Western blot和ELISA方法鉴定其特异性和效价。结果:制备的兔抗人OLFM1多克隆抗体效价为1∶32000,蛋白质印迹证实该抗体具有较好的反应性和特异性。结论:利用生物信息学软件成功地预测出OLFM1蛋白的抗原性,并制备了效价较高的特异性抗人OLFM1多克隆抗体。

Objective：To analyze OLFM1 by bioinformatics and prepare its polyclonal antibody.Methods：According to the bioinformatics analysis and prediction of the possible high structure,hydrophobicity and antigenicity of OLFM1 and the principal of antibody project,a partial peptide of OLFM1 with 20-amino acid residues with was synthesized after homology search.The synthesized peptide was then used to immunize rabbit.The specificity and titer of polyclonal antibody against OLFM1 were identified via ELISA and Western blot.Results：Polyclonal antibody against OLFM1 was proved to recognize OLFM1 protein specifically,and its titers reached 1×32 000.Conclusion：By the bioinformatics analysis and prediction,the hydrophilicity and antigenicity of OLFM1 are analyzed.The polyclonal antibody against OLFM1 is successfully obtained.