Protein microarrays for highly parallel detection and quantitation of specific proteins and antibodies in complex solutionsReport as inadecuate




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Genome Biology

, 2:research0004.1

First Online: 22 January 2001Received: 09 November 2000Revised: 05 December 2000Accepted: 13 December 2000

Abstract

BackgroundWe have developed and tested a method for printing protein microarrays and using these microarrays in a comparative fluorescence assay to measure the abundance of many specific proteins in complex solutions. A robotic device was used to print hundreds of specific antibody or antigen solutions in an array on the surface of derivatized microscope slides. Two complex protein samples, one serving as a standard for comparative quantitation, the other representing an experimental sample in which the protein quantities were to be measured, were labeled by covalent attachment of spectrally resolvable fluorescent dyes.

ResultsSpecific antibody-antigen interactions localized specific components of the complex mixtures to defined cognate spots in the array, where the relative intensity of the fluorescent signal representing the experimental sample and the reference standard provided a measure of each protein-s abundance in the experimental sample. To test the specificity, sensitivity and accuracy of this assay, we analyzed the performance of 115 antibody-antigen pairs. 50% of the arrayed antigens and 20% of the arrayed antibodies provided specific and accurate measurements of their cognate ligands at or below concentrations of 0.34 μg-ml and 1.6 μg-ml, respectively. Some of the antibody-antigen pairs allowed detection of the cognate ligands at absolute concentrations below 1 ng-ml, and partial concentrations of 1 part in 10, sensitivities sufficient for measurement of many clinically important proteins in patient blood samples.

ConclusionsThese results suggest that protein microarrays can provide a practical means to characterize patterns of variation in hundreds of thousands of different proteins in clinical or research applications.

Electronic supplementary materialThe online version of this article doi:10.1186-gb-2001-2-2-research0004 contains supplementary material, which is available to authorized users.

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Author: Brian B Haab - Maitreya J Dunham - Patrick O Brown

Source: https://link.springer.com/



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