ELISA法分析人甲状旁腺激素的抗原表位分布

目的分析人甲状旁腺激素(hPTH)的抗原表位分布,为制备表位特异性抗体和建立hPTH测定方法提供理论依据,为研究hPTH结构和生理功能间的关系提供理论基础。方法采用酶联免疫吸附测定法(ELISA),研究多肽片段hPTH(1-4)、(1-10)、(1-34)、(1-37)、(13-27)、(24-37)、(44-68)、(54-84)及(1-84)与抗全段hPTH(1-84)和抗N端hPTH(1-37)抗体的反应,抗多肽片段hPTH(1-37)、(44-68)、(54-84)及(1-84)抗体与完整hPTH(1-84)抗原的反应,对hPTH尤其是其N端肽抗原表位进行分析。结果hPTH(1-34)、(1-37)、(13-27)与抗hPTH(1-84)及抗hPTH(1-37)抗体,hPTH(44-68)、(54-84)与抗hPTH(1-84)抗体,hPTH(24-37)与抗hPTH(1-37)抗体,以及抗hPTH(54-84)、抗hPTH(44-68)、抗hPTH(1-37)抗体与hPTH(1-84)之间均有较好结合。hPTH(1-4)、(1-10)与抗hPTH(1-37)抗体也呈现出结合梯度。结论在hPTH(1-4)、(1-10)、(1-34)、(1-37)、(13-27)、(24-37)、(44-68)和(54-84)内均存在抗原表位,其中hPTH(13-27)和(24-37)区域是制备特异抗体建立全段或N端hPTH检测方法的两个有价值的抗原表位。

Analyzing the epitopes in human parathyroid hormone by ELISA method

Objective To analyze the epitopes in human parathyroid hormone(hPTH) for producing hPTH antibodies and studying the correlation between structure and function of hPTH.Methods The epitopes in hPTH(1-84) were measured by ELISA method.The interactions were among some PTH peptides interacting with hPTH(1-84) or(1-37) antibody,and among PTH(1-84) interacting with some PTH peptide antibodies.Results Immobilized hPTH(134),(1-37),(13-27) could recognize both hPTH(1-84) antibodies and hPTH(1-37) antibodies;immobilized hPTH(4468) and(54-84) could recognize hPTH(1-84) antibodies;immobilized hPTH(24-37) could recognize hPTH(1-37) antibodies,and the ability of immobilized hPTH(1-4) and(1-10) to combine with hPTH(1-37) antibodies were weak;immobilized hPTH(1-84) could recognize antibodies raised by hPTH(54-84),hPTH(44-68),and hPTH (1-37).Conclusion There are epitopes in the regions of hPTH(1-4),(1-10),(1-34),(1-37),(13-27),(24-37),(44-68),and(54-84).And the regions of hPTH(13-27) and(24-37) may be two valuable epitopes for producing antibodies.