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

doi:10.3969/j.issn.1674-5817.2013.06.004

Abstract

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

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.

References

[1] Joong-Ho W, Ofir G, Shai SS, et al.Significance analysis of xMap cytokine bead arrays[J]. Proc Natl Acad Sci USA, 2012, 109(8):2848-2853.
[2] Elizabeth CB, Sandra MR, Christopher C, et al.Multisite Comparison of High-Sensitivity Multiplex Cytokine Assays[J]. Clin Vaccine Immunol, 2011, 18(8):1229-1242.
[3] Aleksandra IK, Andrea N, Jacqueline M, et al.Comparison of ophthalmic sponges and extraction buffers for quantifying cytokine profiles in tears using Luminex technology[J]. Mol Vis, 2012, 18: 2717-2725.
[4] Giavedoni LD.Simultaneous detection of multiple cytokines and chemokines from nonhuman primates using luminex technology[J]. J Immunol Methods, 2005, 301(1-2):89-101.
[5] Datta SC, Opp MR.Lipopolysaccharide-induced increases in cytokines in discrete mouse brain regions are detectable using Luminex xMAP technology[J]. J Neurosci Methods, 2008, 175(1):119-124.
[6] Brett H.Bio-Rad’s Bio-Plex??suspension array system, xMAP technology overview[J]. Arch Physiol Biochem, 2012, 118(4):192-196.
[7] Paulo JA, Lee LS, Wu B,et al.Cytokine profiling of pancreatic fluid using the ePFT collection method in tandem with a multiplexed microarray assay[J]. J Immunol Methods, 2011, 369(1-2):98-107.
[8] Djoba Siawaya JF, Roberts T, Babb C, et al.An evaluation of commercial fluorescent bead-based luminex cytokine assays[J]. PLoS One, 2008, 3(7):e25-35.
[9] Jung S, Oh EJ, Yang CW, et al.Comparative evaluation of ELISA and Luminex panel reactive antibody assays for HLA alloantibody screening[J]. Korean J Lab Med, 2009, 29(5):473-480.
[10] Wang LS, Leung YY, Chang SK, et al.Comparison of xMAP and ELISA assays for detecting cerebrospinal fluid biomarkers of Alzheimer’s disease[J]. J Alzheimers Dis, 2012, 31(2): 439-445.
[11] Codorean E, Nichita C, Albulescu L, et al.Correlation of XMAP and ELISA cytokine profiles; development and validation for immunotoxicological studies in vitro[J]. Roum Arch Microbiol Immunol, 2010, 69(1):13-19.
[12] Li C, Song Y, Luan S, et al.Research on the stability of a rabbit dry eye model induced by topical application of the preservative benzalkonium chloride[J]. PLoS One, 2012, 7(3):e33688.
[13] Luo WY, Lin ZX, Li H, et al.Chronic toxicity of methylamine on cardiovascular endothelium of rabbits[J]. Chin J Pharmacol Toxicol, 2008, 22(1):24-30.
[14] Mader A, Kunert R.Evaluation of the potency of the anti-idiotypic antibody Ab2/3H6 mimicking gp41 as an HIV-1 vaccine in a rabbit prime/boost study[J]. PLoS One, 2012, 7(6):e39063.
[15] 陈方明, 陈民利, 陈亮, 等. WHBE兔脾虚型肠易激综合征模型舌象病理组织形态学观察[J]. 中国比较医学杂志, 2009, 19(8):15-18.
[16] 应华忠, 寿旗扬, 陈民利, 等. WHBE兔的血液学指标测定与比较[J]. 实验动物与比较医学, 2010, 30(1):44-47.
[17] 陈民利, 蔡月琴, 陶涛, 等. 微卫星标记对WHBE兔封闭群、日本大耳白兔和新西兰兔的遗传分析[J]. 中国比较医学杂志, 2008, 18(9):21-27.
[18] 蔡月琴, 屠珏, 余佳, 等. RAPD标记技术用于WHBE兔近交系培育中的遗传分析[J]. 中国实验动物学报, 2009, 17(5): 326-329.