碱性磷酸酶标记抗人绒毛膜促性腺激素抗体的工艺比较

Process comparison of conjugating alkaline phosphatase with anti-human chorionic gonadotropin antibody

目的通过测定最低检测限、精密性、热稳定性和临床相关性来比较3种碱性磷酸酶(alkaline phosphatase,AP)标记抗人绒毛膜促性腺激素(human chorionic gonadotropin,HCG)抗体工艺的优劣。方法用3种常用的蛋白质标记方法,将碱性磷酸酶和抗HCG的单抗进行偶联,进一步制成HCG的化学发光法定量测定试剂盒。结果对用3种标记工艺获得的试剂的各项性能指标进行考察,结果过碘酸钠法各指标表现均衡,测值准确;4-马来酰亚胺基丁酸-N-琥珀酰亚胺酯[N-(4-Maleimidobutyryloxy)succinimide,GMBS]法灵敏度高但热稳定性不佳;戊二醛法灵敏度过低且样本比对结果偏高。结论用过碘酸钠法标记碱性磷酸酶与单抗获得的试剂在最低检测限、精密性、热稳定性及样本测定值比对各项指标中表现良好,是本研究推荐的一种标记工艺。

Objective To compare 3 conjugating processes of alkaline phosphatase（AP） with anti-human chorionic go- nadotropin（HCG） monoclonal antibody by determing the limit of detection, precision, thermal accelerated stability and clinical rele- vance. Methods AP and anti-HCG McAb were conjugated by employing 3 commonly used protein-coupled methods. Further the HCG quantitative detection kit was developed. Results All the performance characteristics of the 3 conjugating processes were compared. The sodium periodate method performed best with accurate measurement; the 4-maleimidobutyric acid N-hydroxysuccinimide （GMBS） method performed with high precision but bad thermal accelerated stability; and the glutaral performed with low precision and poor clinical relevance. Conclusion The sodium periodate conjugating method was recommended by this study for it＇ s well performing in the the limit of detection, precision, thermal accelerated stability and clinical relevance.