兔免疫球蛋白液相芯片的制备及其检测方法的建立

doi:10.3969/j.issn.1674-5817.2013.06.004

实验动物与比较医学 ›› 2013, Vol. 33 ›› Issue (6): 433-438.DOI: 10.3969/j.issn.1674-5817.2013.06.004

兔免疫球蛋白液相芯片的制备及其检测方法的建立

蔡月琴, 徐剑钦, 徐孝平, 王德军, 黄宇, 吴蔚, 陈民利

浙江中医药大学动物实验研究中心/比较医学研究中心, 杭州 310053

收稿日期:2013-05-07 出版日期:2013-12-25 发布日期:2013-12-25
作者简介:蔡月琴(1982-), 女, 助理研究员, 研究方向: 分子生物学与免疫学。E-mail: cyq810@hotmail.com
基金资助:
1. 浙江省教育厅重大科技攻关项目(ZD2009004); 2. 浙江省科技计划项目(2011C37093); 3. “浙江省卫生高层次创新人才培养工程项目”资助

Development of Liquid Suspension Chip of Rabbit Immune Globulins and Flexible Multi-analyte Profiling

CAI Yue-qin, XU Jian-qin, XU Xiao-ping, WANG De-jun, HUANG Yu, WU Wei, CHEN Min-li

Laboratory Animal Research Center, Zhejiang Chinese Medical University , Hangzhou 310053, China

Received:2013-05-07 Online:2013-12-25 Published:2013-12-25

摘要/Abstract

摘要： 目的制备实验兔的免疫球蛋白液相芯片, 建立液相芯片技术(xMAP)检测方法。方法分别将兔IgG、IgM和IgA亲和纯化的捕获抗体耦联至荧光编码微球,对蛋白耦联量进行优化,制备兔IgG、IgM和IgA液相芯片,同步检测白毛黑眼兔(WHBE兔)和日本大耳白兔(JW兔)血清中IgG、IgM和IgA相对含量,建立可应用于实验兔免疫球蛋白半定量检测的xMAP方法,并对其结果用ELISA方法进行验证。结果 (1)IgG、IgM和IgA的最佳蛋白耦联量分别为10、20和15 μg。(2)xMAP检测结果显示,WHBE兔血清中IgG、IgM蛋白含量显著低于JW兔(P<0.05),IgA蛋白含量显著高于JW兔(P<0.01); ELISA检测结果表明,WHBE兔血清中IgG、IgM蛋白含量显著低于JW兔(P<0.05,P<0.01),其中IgM蛋白含量变化达到极显著水平,IgA蛋白含量显著高于JW兔(P<0.01)。结论应用制备的兔IgG、IgM和IgA液相芯片建立的xMAP方法,对实验兔血清进行检测,检测结果与ELISA结果一致,提示该方法可应用于实验兔血清的IgG、IgM和IgA半定量检测。

关键词: 免疫球蛋白, 液相芯片, xMAP, 实验兔

Abstract: Objective To establish flexible multi-analyte profiling (xMAP) in rabbit immune globulins using the Luminex liquid suspension array system. Methods The rabbit IgG, IgM and IgA affinity capture antibody was conjugated to different fluorescent encoded microspheres, respectively. The conjuctional capacity of rabbit IgG, IgM and IgA coupling antibody was optimized, and the liquid suspension chip of rabbit IgG, IgM and IgA was developed. The simultaneous measurement of multiple immune globulins in WHBE and JW rabbit was semi-quantitative detected by xMAP in liquid suspension array system. And verify the rabbit immune globulins results by ELISA analysis. Results (1) The optimal conjuctional capacity of rabbit IgG, IgM and IgA coupling antibody was 10 μg, 20 μg and 15 μg, respectively. (2) IgG, IgM and IgA in WHBE rabbit and JW rabbit were simultaneously detected by xMAP technology. IgG and IgM in WHBE rabbit were significantly lower than JW rabbit (P<0.05), and IgA in WHBE rabbit was significantly higher than JW rabbit (P<0.01). Meanwhile, IgG, IgM and IgA in WHBE rabbit and JW rabbit was respectively analyzed by ELISA method. IgG and IgM in WHBE rabbit were significantly lower than JW rabbit (P<0.05, P<0.01), and IgA in WHBE rabbit was significantly higher than JW rabbit (P<0.01). Conclusion The xMAP for semi-quantitative detecting rabbit immune globulins has been successfully established. And all the results were consistent with the ELISA results.

Key words: Immune globulins, Liquid suspension chip, xMAP, Rabbit

蔡月琴, 徐剑钦, 徐孝平, 王德军, 黄宇, 吴蔚, 陈民利. 兔免疫球蛋白液相芯片的制备及其检测方法的建立[J]. 实验动物与比较医学, 2013, 33(6): 433-438.

CAI Yue-qin, XU Jian-qin, XU Xiao-ping, WANG De-jun, HUANG Yu, WU Wei, CHEN Min-li. Development of Liquid Suspension Chip of Rabbit Immune Globulins and Flexible Multi-analyte Profiling[J]. Laboratory Animal and Comparative Medicine, 2013, 33(6): 433-438.

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兔免疫球蛋白液相芯片的制备及其检测方法的建立

Development of Liquid Suspension Chip of Rabbit Immune Globulins and Flexible Multi-analyte Profiling

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