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

作　　者：靳颖[1] 宋晓娇 王静 徐亮[4] 袁婳婳 王冬梅[4] JIN Ying;SONG Xiao-jiao;WANG Jing;XU Liang;YUAN Hua-hua;WANG Dong-mei(Department of Clinical Laboratory,Affiliated Hospital of Logistics University of PAP,Tianjin 300162,China)

机构地区：[1]武警后勤学院附属医院检验科,天津300162 [2]天津药业研究院有限公司,天津300457 [3]天津市工业化学研究所有限公司,天津300042 [4]天津市临床药物关键技术重点实验室,天津医科大学药学院,天津300070

出　　处：《武警后勤学院学报（医学版）》2018年第1期15-18,共4页Journal of Logistics University of PAP（Medical Sciences）

基　　金：武警后勤学院基础研究项目(WHJ2014017);天津医科大学大学生学术研究资助计划项目(TMUUROP2015-19);天津市科技计划项目(15YFFCTG00130)

摘　　要：【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.

关 键 词：羧化聚苯乙烯微球 胶乳增强免疫比浊法 Β2微球蛋白 影响因素 

分 类 号：R943[医药卫生—药剂学]