【摘要】 建立多抗夹心ELISA测定人血浆普莱单抗(anti-GPIIb/IIaF(ab)2)浓度的方法并进行方法学验证。采用复合快速免疫法免疫家兔获得抗血清,经常规及普莱单抗特异性亲和层析纯化,并以人AB型血浆偶联的Sephorose 4B凝胶去除与人血浆的交叉反应。以获得的特异性多抗包板,生物素标记的该多抗为检测抗体,建立测定人血浆普莱单抗浓度的多抗夹心ELISA法,并进行了方法学验证。复合快速免疫法可在较短的时间内获得了高滴度的抗普莱单抗的多抗;利用所获得的多抗构建的ELISA试剂盒测定普莱单抗的浓度范围为1.56~50ng·mL-1,方法学认证的各项指标符合药代动力学研究的要求,为进行临窗前及临床普莱单抗药物代谢动力学研究奠定了坚实的基础,为其它单抗及高分子生物技术药物的药代动力学研究提供了借鉴思路。更多还原

【Abstract】 An enzyme-linked immunosorbent assays (ELISA) for measurement the concentration of Pulaimab (anti-GPIIb/IIIa mAB F(ab)2) in human plasma was developed. Pulaimab was used in order to raise polyclonal anti-Pulaimab antibodies in rabbits through a complex immunology method. Antisera was purified with Protein G, Pulaimab coupled Sephrose 4B and Human AB blood type plasma coupled Sephrose 4B affinity chromatography, respectively. Antisera was achieved in one month with high titer. A sandwich ELISA kit was set up using the purified polyclonal antibodies to measure Pulaimab concentration in plasma. The assay allowed the detection of Pulaimab in the quantification range 1.56～50ng·mL-1, in biomatrix. The method is a rapid and simple immunoassay for monitoring of Pulaimab in preclinical and clinical use, and it can be used to kinetics study of protein polypeptide drugs.更多还原