蛋白芯片制作与应用（4）－液态芯片

液态芯片原理

编码微球：分别用不同配比的两种荧光染料将直径5.6μm的聚苯乙烯微球（Beads）染成不同的荧光色，从而获得多达100种经荧光编码的微球。

交联探针、抗体或抗原：把针对不同检测物的核酸探针、抗体或抗原以共价方式结合到特定荧光编码的微球上。


检测反应：先把针对不同检测物的、用不同荧光色编码的微球混合，再加入被检测物（可以是血清中的抗原、抗体或酶等，也可以是PCR产物）。 悬液中的微球与被检测物特异性结合，结合物被标记上荧光物质。


激光分析：微球成单列通过两束激光，一束判定微球的荧光编码；另一束测定微球上的报告分子的荧光强度。

1）液态芯片目前仍然处于基础研究方面，虽然已经有部分临床研究，但可能离大规模应用还有一段距离。
2）常见的应用如肿瘤、内分泌、自身免疫等的检测，也有人将其用于细胞因子谱判断。
3）下面是一些液态芯片相关经典的文献，有时间参考一下
1. Hany Ezzeldin, Yoshihiro Okamoto, Martin R. Johnson, et al. A High-qhroughput Denaturing High - Performance Liquid Chromatography Method for the Identification of Variant Alleles Associated with Dihydropyrimidine Dehydrogenase Deficiency. Analytical Biochemistry,2002,306:63 - 73..
2. Carson RT, Vignali DA. Simultaneous quantitation of 15 cytokines using a multiplexed flow cytometric assay. J Immunol Methods, 1999,30.227(1 -2) :41 -52..
3. Karlar,G. A, M. Jeddi-Tehrani, F. Shokri. Diminished Thl and Th2 cytokine production in healthy adult nonresponders to recombinant hepatitis B vaccine. Scand. J Immunol,2002 ,55 :311-314..
4. Taylor JD,Briley D ,Nguyen Q,et al. Flow cytometric platform for high throughput single nucleotide polymorphism analysis. Biotechniques,2001.30 ( 3 ) ,661 - 666.668 - 669..
5. Wilco de Jager, Henk te Velthuis, Berent J. Prakken, et al. Simultaneous Detection of 15 Human Cytokines in a Single Sample of Stimulated Peripheral Blood Mononuclear Ceils. Clinical and Diagnostic Laboratory Immunology,2003,10( 1 ) :133-139..
6. lannone MA.Microsphere-htsed molecular cytometry. Clin Lab Med,2001.21 (4) :731 -742..
7. Joos TO, Stoll D, Tempiin MF. Miniaturised multiplexed immunoassays. Curr Opin Chem Biol,2002,6( 1 ) :76 -80..
8. Dunbar SA, Vander zee CA, Oliver KG, et al. Quantitative, multiplexed detection of bacterial pathogens : DNA and protein applications of the Luminex Lab MAP system. J Microbiol Methods,2003,53 (2) :245 - 252..
9. David Opalka, Charles E. Lachman, Stefani A. MacMullen, et al. Simuhaneous Quantitation of Antibodies to Neutralizing Epitopes on Virus-Like Particles for Human Papillomavirus Types 6,11,16, and 18 by a Multiplexed Luminex Assay. Clinical and Diagnostic Laboratory Immunology,2003,10( 1 ) :108-115..