The CombiMatrix antibody microarray is a versatile, sensitive detection platform based on the generation and transduction of electrochemical signals following antigen binding to surface antibodies. The sensor chip described herein is comprised of microelectrodes coupled to an adjacent bio-friendly matrix coated with antibodies to the biological pathogens Yersinia pestis and Bacillus anthracis, and the bacterial toxin staphylococcal enterotoxin B (SEB). Using this system, we were able to detect SEB and inactivated Y. pestis individually as well as in two-plex assays at concentrations as low as 5 pg/mL and 106 CFU/mL, respectively. We also introduce super avidin-biotin system (SABS) as a viable and effective means to enhance assay signal responses and lower detection limits. Together these technologies represent substantial advances in point-of-care and point-of-use detection applications.
References
[1]
Lipshutz, R.J.; Fodor, S.P.; Gingeras, T.R.; Lockhart, D.J. High density synthetic oligonucleotide arrays. Nat. Genet？1999, 21, 20–24, doi:10.1038/4447. 9915496
[2]
Wilgenbus, K.K.; Lichter, P. DNA chip technology ante portas. J. Mol. Med？1999, 77, 761–768, doi:10.1007/s001099900057. 10619436
[3]
Hacia, J.G.; Brody, L.C.; Collins, F.S. Applications of DNA chips for genomic analysis. Mol. Psychiat？1998, 3, 483–492, doi:10.1038/sj.mp.4000475.
Gibson, G. The environmental contribution to gene expression profiles. Nat. Rev. Genet？2008, 9, 575–581. 18574472
[6]
Yoo, S.M.; Lee, S.Y. Diagnosis of pathogens using DNA microarray. Recent. Pat. Biotechnol？2008, 2, 124–129, doi:10.2174/187220808784619711. 19075860
[7]
MacBeath, G. Protein microarrays and proteomics. Nat. Genet？2002, 32, 526–532, doi:10.1038/ng1037. 12454649
[8]
Stoll, D.; Templin, M.F.; Schrenk, M.; Traub, P.C.; Vohringer, C.F.; Joos, T.O. Protein microarray technology. Front. Biosci？2002, 7, 13–32, doi:10.2741/stoll.
[9]
Templin, M.F.; Stoll, D.; Schrenk, M.; Traub, P.C.; Vohringer, C.F.; Joos, T.O. Protein microarray technology. Trend. Biotechnol？2002, 20, 160–166, doi:10.1016/S0167-7799(01)01910-2.
[10]
Wolf-Yadlin, A.; Sevecka, M.; MacBeath, G. Dissecting protein function and signaling using protein microarrays. Curr. Opin. Chem. Biol？2009, 13, 398–405, doi:10.1016/j.cbpa.2009.06.027. 19660979
[11]
Stoevesandt, O.; Taussig, M.J.; He, M. Protein microarrays: high-throughput tools for proteomics. Expert Rev. Proteomics？2009, 6, 145–157, doi:10.1586/epr.09.2. 19385942
Dill, K.; Montgomery, D.; Wang, W.; Tsai, J. Antigen detection using microelectrode array microchips. Analytica chimica Acta？2001, 444, 69–78, doi:10.1016/S0003-2670(01)01155-2.
[14]
Dill, K.; Montgomery, D.D.; Ghindilis, A.L.; Schwarzkopf, K.R.; Ragsdale, S.R.; Oleinikov, A.V. Immunoassays based on electrochemical detection using microelectrode arrays. Biosens. Bioelectron？2004, 20, 736–742, doi:10.1016/j.bios.2004.06.049. 15522588
[15]
Afshari, C.A.; Nuwaysir, E.F.; Barrett, J.C. Application of complementary DNA microarray technology to carcinogen identification, toxicology, and drug safety evaluation. Cancer Res？1999, 59, 4759–4760. 10519378
[16]
Byrne, B.; Stack, E.; Gilmartin, N.; O’Kennedy, R. Antibody-Based Sensors: Principles, Problems and Potential for Detection of Pathogens and Associated Toxins. Sensors？2009, 9, 4407–4445, doi:10.3390/s90604407. 22408533
[17]
Rowe, C.A.; Scruggs, S.B.; Feldstein, M.J.; Golden, J.P.; Ligler, F.S. An array immunosensor for simultaneous detection of clinical analytes. Anal. Chem？1999, 71, 433–439, doi:10.1021/ac980798t. 9949731
[18]
Poli, M.A.; Rivera, V.R.; Neal, D. Sensitive and specific colorimetric ELISAs for Staphylococcus aureus enterotoxins A and B in urine and buffer. Toxicon？2002, 40, 1723–1726, doi:10.1016/S0041-0101(02)00202-7. 12457885
[19]
Homola, J.; Dostalek, J.; Chen, S.; Rasooly, A.; Jiang, S.; Yee, S.S. Spectral surface plasmon resonance biosensor for detection of staphylococcal enterotoxin B in milk. Int. J. Food Microbiol？2002, 75, 61–69, doi:10.1016/S0168-1605(02)00010-7. 11999118
[20]
Anderson, G.P.; Taitt, C.R. Amplification of microsphere-based microarrays using catalyzed reporter deposition. Biosens. Bioelectron？2008, 24, 324–328, doi:10.1016/j.bios.2008.03.045. 18485692
[21]
Tempelman, L.A.; King, K.D.; Anderson, G.P.; Ligler, F.S. Quantitating staphylococcal enterotoxin B in diverse media using a portable fiber-optic biosensor. Anal Biochem？1996, 233, 50–57, doi:10.1006/abio.1996.0006. 8789146
[22]
Shriver-Lake, L.C.; Shubin, Y.S.; Ligler, F.S. Detection of staphylococcal enterotoxin B in spiked food samples. J. Food Prot？2003, 66, 1851–1856. 14572223
[23]
Sapsford, K.E.; Taitt, C.R.; Loo, N.; Ligler, F.S. Biosensor detection of botulinum toxoid A and staphylococcal enterotoxin B in food. Appl. Environ. Microbiol？2005, 71, 5590–5592, doi:10.1128/AEM.71.9.5590-5592.2005. 16151154
