Suspension microarray immunoassay signal amplification using multilayer formation

被引:12
作者
Anderson, George P. [1 ]
Taitt, Chris R. [1 ]
机构
[1] USN, Res Lab, Ctr Biomol Sci & Engn, Washington, DC 20375 USA
关键词
Luminex xMAP; microarray; immunoassay; avidin-biotin; anti-streptavidin antibody; super avidin-biotin system; fluid array immunoassay; flow Cytometry; microsphere;
D O I
10.1166/sl.2008.018
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
A method for rapid amplification of signals is demonstrated in multiplexed fluid microarrays using the 'super avidin-biotin system' (SABS). The Luminex xMAP system is a specialized flow cytometer that performs multiplexed microsphere based fluoroimmunoassays. The xMAP system's standard protocol utilizes antibodies immobilized to microspheres as the capture surface for target antigens and complementary biotinylated antibodies serve as the recognition "tracer" reagents. This antibody:target:biotinylated antibody complex is fluorescently labeled by the addition of streptaviclin-phycoerythrin (SA-PE). The present study shows that addition of two simple steps greatly amplifies the signal produced by the standard protocol, typically by a factor of similar to 10. After addition of the SA-PE, the microspheres are subsequently exposed to biotinylated goat anti-streptavidin antibody; this is then followed by a second round of SA-PE, which further amplifies the initial signal. This amplification protocol was demonstrated for multiplexed detection of cholera toxin and botulinum toxoid A. In addition, several SA-PE conjugates having a variety of SA-to-PE stoichiometries were evaluated, as well as an alternative protocol, wherein biotinylated PE was substituted for the anti-streptavidin antibody.
引用
收藏
页码:213 / 218
页数:6
相关论文
共 17 条
[1]   Development of a luminex based competitive immunoassay for 2,4,6-trinitrotoluene (TNT) [J].
Anderson, George P. ;
Lamar, Jacqueline D. ;
Charles, Paul T. .
ENVIRONMENTAL SCIENCE & TECHNOLOGY, 2007, 41 (08) :2888-2893
[2]   Suspension microarray with dendrimer signal amplification allows direct and high-throughput subtyping of Listeria monocytogenes from genomic DNA [J].
Borucki, MK ;
Reynolds, J ;
Call, DR ;
Ward, TJ ;
Page, B ;
Kadushin, J .
JOURNAL OF CLINICAL MICROBIOLOGY, 2005, 43 (07) :3255-3259
[3]   DETECTION OF YERSINIA-PESTIS FRACTION-1 ANTIGEN WITH A FIBER OPTIC BIOSENSOR [J].
CAO, LK ;
ANDERSON, GP ;
LIGLER, FS ;
EZZELL, J .
JOURNAL OF CLINICAL MICROBIOLOGY, 1995, 33 (02) :336-341
[4]  
CHEN A, 2007, PLAN XMAP S 2007 ABS
[5]   ENUMERATION OF CR-1 COMPLEMENT RECEPTORS ON ERYTHROCYTES USING A NEW METHOD FOR DETECTING LOW-DENSITY CELL-SURFACE ANTIGENS BY FLOW-CYTOMETRY [J].
COHEN, JHM ;
AUBRY, JP ;
JOUVIN, MH ;
WIJDENES, J ;
BANCHERAU, J ;
KAZATCHKINE, M ;
REVILLARD, JP .
JOURNAL OF IMMUNOLOGICAL METHODS, 1987, 99 (01) :53-58
[6]   Multiplex assay for simultaneous measurement of antibodies to multiple Plasmodium falciparum antigens [J].
Fouda, Genevieve G. ;
Leke, Rose F. G. ;
Long, Carole ;
Druilhe, Pierre ;
Zhou, Ainong ;
Taylor, Diane Wallace ;
Johnson, Armead H. .
CLINICAL AND VACCINE IMMUNOLOGY, 2006, 13 (12) :1307-1313
[7]   Comparison of electrochemiluminescence assay and ELISA for the detection of Clostridium botulinum type B neurotoxin [J].
Guglielmo-Viret, V ;
Attrée, O ;
Blanco-Gros, V ;
Thullier, P .
JOURNAL OF IMMUNOLOGICAL METHODS, 2005, 301 (1-2) :164-172
[8]   Sensitive and specific detection of staphylococcal epidermolysins A and B in broth cultures by flow cytometry-assisted multiplex immunoassay [J].
Joubert, O ;
Keller, D ;
Pinck, A ;
Monteil, H ;
Prévost, G .
JOURNAL OF CLINICAL MICROBIOLOGY, 2005, 43 (03) :1076-1080
[9]   IMMUNOCHEMICAL DETECTION OF PROTEINS BIOTINYLATED ON NITROCELLULOSE REPLICAS [J].
LAROCHELLE, WJ ;
FROEHNER, SC .
JOURNAL OF IMMUNOLOGICAL METHODS, 1986, 92 (01) :65-71
[10]   Tris (2,2'-bipyridyl)ruthenium(II) electrogenerated chemiluminescence in analytical science [J].
Lee, WY .
MIKROCHIMICA ACTA, 1997, 127 (1-2) :19-39