Multifunctional protein microarrays for cultivation of cells and immunodetection of secreted cellular products

Caroline N. Jones, Youn Lee Ji, James Zhu, Gulnaz Stybayeva, Erlan Ramanculov, Mark A Zern, Alexander Revzin

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

The microarray format is being used extensively for combinatorial screening of cellular interactions with proteins, small molecules, or biomaterials. The utility of microarray-based cell cultivation approaches may be enhanced further by incorporating biosensing elements alongside the cell-adhesive ligands to enable local detection of secreted cellular products. The concept of combining cells and sensing elements in the same microarray is demonstrated in the present paper with hepatocytes serving as a model cellular system. Robotic microarraying was employed to print arrays of 300-μm-diameter collagen (I) spots alongside the antibody (Ab) spots specific to liver proteins: albumin and α1-antitrypsin (α1-AT). Protein microarrays were printed onto poly(ethylene glycol) hydrogel-coated glass slides, thus eliminating nonspecific adsorption of cells or proteins. When incubated with printed microarrays, hepatocytes became localized on collagen (I) domains but did not attach on Ab spots or elsewhere on hydrogel-coated glass substrates. Liver-specific proteins secreted by hepatocytes were captured on Ab domains in the immediate vicinity of the cells, detected with a sandwich immunofluorescent assay and quantified using a microarray scanner. Importantly, hepatic albumin and α1-AT production detected in the microarray was comparable to enzyme-linked immunosorbent assay measurements of these proteins. In the future, the juxtaposition of sensing Ab regions with cell arrays will be particularly useful for the detection of local appearance or loss of phenotype of cells interacting with the printed components of the cellular microenvironment.

Original languageEnglish (US)
Pages (from-to)6351-6357
Number of pages7
JournalAnalytical Chemistry
Volume80
Issue number16
DOIs
StatePublished - Aug 15 2008

ASJC Scopus subject areas

  • Analytical Chemistry

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    Jones, C. N., Ji, Y. L., Zhu, J., Stybayeva, G., Ramanculov, E., Zern, M. A., & Revzin, A. (2008). Multifunctional protein microarrays for cultivation of cells and immunodetection of secreted cellular products. Analytical Chemistry, 80(16), 6351-6357. https://doi.org/10.1021/ac8007626