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Simultaneous multianalyte ELISA performed on a microarray platform.
Clin Chem. 2001 Aug; 47(8):1451-7.CC

Abstract

BACKGROUND

A logical progression of the widely used microtiter plate ELISA is toward a protein array format that allows simultaneous detection of multiple analytes at multiple array addresses within a single well. Here we describe the construction and use of such a multiplex ELISA to measure prostate-specific antigen (PSA), alpha1-antichymotrypsin-bound PSA (PSA-ACT), and interleukin-6 (IL-6).

METHODS

We silanized glass plates and printed the appropriate capture antibodies to allow for the construction of "sandwich" ELISA quantification assays. We examined specificity of the assay for appropriate antigen, assembled calibration curves, and obtained PSA concentrations for 14 human serum samples. We compared the serum PSA concentrations derived through the use of our array with values obtained independently using a standard ELISA method.

RESULTS

R2 values generated by our microarray for the PSA and PSA-ACT calibration curves were 0.989 and 0.979, respectively. Analyte concentrations used for the construction of these curves were 0.31-20 microg of protein/L of diluent. IL-6 calibration curve concentrations were 4.9-300 ng of IL-6/L of diluent. The R2 value for the IL-6 calibration curve was 0.983. The 14 human serum samples screened by this micro-ELISA technique for PSA concentrations generated a regression equation (linear) with a slope of 0.83 +/- 0.10 and intercept of 0.74 +/- 0.70 (R2 = 0.88).

CONCLUSIONS

Multiplexed ELISA arrays are a feasible option for analyte quantification in complex biologic samples.

Authors+Show Affiliations

Genometrix, Inc., 2700 Research Forest Dr., The Woodlands, TX 77381, USA. rwiese@genometrix.comNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

11468236

Citation

Wiese, R, et al. "Simultaneous Multianalyte ELISA Performed On a Microarray Platform." Clinical Chemistry, vol. 47, no. 8, 2001, pp. 1451-7.
Wiese R, Belosludtsev Y, Powdrill T, et al. Simultaneous multianalyte ELISA performed on a microarray platform. Clin Chem. 2001;47(8):1451-7.
Wiese, R., Belosludtsev, Y., Powdrill, T., Thompson, P., & Hogan, M. (2001). Simultaneous multianalyte ELISA performed on a microarray platform. Clinical Chemistry, 47(8), 1451-7.
Wiese R, et al. Simultaneous Multianalyte ELISA Performed On a Microarray Platform. Clin Chem. 2001;47(8):1451-7. PubMed PMID: 11468236.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Simultaneous multianalyte ELISA performed on a microarray platform. AU - Wiese,R, AU - Belosludtsev,Y, AU - Powdrill,T, AU - Thompson,P, AU - Hogan,M, PY - 2001/7/27/pubmed PY - 2001/8/10/medline PY - 2001/7/27/entrez SP - 1451 EP - 7 JF - Clinical chemistry JO - Clin. Chem. VL - 47 IS - 8 N2 - BACKGROUND: A logical progression of the widely used microtiter plate ELISA is toward a protein array format that allows simultaneous detection of multiple analytes at multiple array addresses within a single well. Here we describe the construction and use of such a multiplex ELISA to measure prostate-specific antigen (PSA), alpha1-antichymotrypsin-bound PSA (PSA-ACT), and interleukin-6 (IL-6). METHODS: We silanized glass plates and printed the appropriate capture antibodies to allow for the construction of "sandwich" ELISA quantification assays. We examined specificity of the assay for appropriate antigen, assembled calibration curves, and obtained PSA concentrations for 14 human serum samples. We compared the serum PSA concentrations derived through the use of our array with values obtained independently using a standard ELISA method. RESULTS: R2 values generated by our microarray for the PSA and PSA-ACT calibration curves were 0.989 and 0.979, respectively. Analyte concentrations used for the construction of these curves were 0.31-20 microg of protein/L of diluent. IL-6 calibration curve concentrations were 4.9-300 ng of IL-6/L of diluent. The R2 value for the IL-6 calibration curve was 0.983. The 14 human serum samples screened by this micro-ELISA technique for PSA concentrations generated a regression equation (linear) with a slope of 0.83 +/- 0.10 and intercept of 0.74 +/- 0.70 (R2 = 0.88). CONCLUSIONS: Multiplexed ELISA arrays are a feasible option for analyte quantification in complex biologic samples. SN - 0009-9147 UR - https://www.unboundmedicine.com/medline/citation/11468236/Simultaneous_multianalyte_ELISA_performed_on_a_microarray_platform_ L2 - http://ovidsp.ovid.com/ovidweb.cgi?T=JS&PAGE=linkout&SEARCH=11468236.ui DB - PRIME DP - Unbound Medicine ER -