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目的制备抗人β2微球蛋白(β2-microglobulin,β2-MG)特异性单克隆抗体并建立其双抗体夹心定量ELISA免疫检测方法。方法以高纯度的人β2-MG作为免疫原,采用常规免疫和脾内免疫相结合的方法免疫纯系Balb/c小鼠,通过杂交瘤技术制备抗人β2-MG单克隆抗体并鉴定,以此为基础建立双抗体夹心ELISA检测方法。结果制备出5株稳定分泌抗人β2-MG的单克隆抗体,经鉴定5株单抗皆为IgG1类,均为β2-MG抗原特异性抗体。经抗体配对试验筛选出1对可用于ELISA检测的配对抗体,建立了定量ELISA检测方法。结论制备出抗人β2-MG单克隆抗体并建立了双抗体夹心定量ELISA免疫检测方法,与国外试剂盒检测结果相关性良好。
Objective To prepare a specific monoclonal antibody against β2-microglobulin (β2-MG) and to establish a double-antibody sandwich immunoassay for its detection. Methods Pure human Balb / c mice were immunized with high-purity human β2-MG and immunized with conventional immunization and splenic immunization. Monoclonal antibodies against human β2-MG were prepared by hybridoma technique and identified by This is based on the establishment of double antibody sandwich ELISA detection method. Results Five monoclonal antibodies against human β2-MG were successfully prepared. All the five monoclonal antibodies were identified as IgG1, all of which were β2-MG antigen-specific antibodies. A pair of matched antibodies that can be used for ELISA detection was screened by antibody pairing test, and a quantitative ELISA detection method was established. Conclusion The anti-human β2-MG monoclonal antibody was prepared and a double antibody sandwich quantitative ELISA immunoassay was established. It has good correlation with the test results of foreign kits.