β2微球蛋白胶乳增强免疫比浊试剂制备的影响因素

Study on the influencing factors in the preparation for the reagents of β2 microglobin latexenhanced turbidimetric immunoassay

【目的】以β2微球蛋白(β2 microglobin,β2-M)为目标物,研究胶乳增强免疫比浊试剂(latex-enhanced turbidimetric immunoassay,LETIA)的制备条件对其检测性能的影响。【方法】采用碳二亚胺法将β2-M抗体偶联到羧基化聚苯乙烯微球(carboxylated polystyrene,PS)的表面,制备了β2-M-LETIA检测试剂。考察胶乳微球的粒径、偶联试剂的用量以及抗体加入量等条件对β2-M-LETIA试剂检测性能的影响。【结果】粒径为120 nm和150 nm的微球具有较好的效果;抗体偶联量随1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)和N-羟基琥珀酰亚胺(N-hydroxysuccinimide,NHS)两种偶联试剂用量的增加而增大,当其用量达到0.25 mg/ml球时抗体的偶联量达到最大值;最佳的抗体加入量为6 mg/ml。【结论】得到了作为载体的微球粒径、EDC和NHS的用量以及抗体的加入量等制备条件对β2-M-LETIA检测试剂的抗体偶联量和检测性能的影响规律。

【Objective】To study the influencing factors in the preparation for the reagents of β2 microglobin（β2-M） latex-enhanced turbidimetric immunoassay（LETIA）.【Methods】The detection reagent of LETIA for β2-M was prepared by coupling β2-M antibodies on the surface of carboxylated polystyrene（PS） nanoparticles with carbodiimide method. The effects of particle size, coupling reagent volume and antibody dosage were explored on performance of β2-M-LETIA.【Results】The microspheres with the diameter of 120 nm and 150 nm were of better effect; the amount of coupling antibodies increased with the dosage of 1-ethyl-3-（3-dimethylaminepropyl）carbodiimidehy-drochloride（EDC） and N-hydroxysuccinimide（NHS） and reached the maximum at 0.25 mg/ml; the optimum amount of antibodies was 6 mg/ml for the coupling.【Conclusion】The influencing tendency of preparation conditions mentioned above is related with the amount of coupling antibodies and detection performance of β2-M-LETIA reagent.