Evaluation of homo- and hetero-functionally activated glass surfaces for optimized antibody arrays

Antibody arrays hold great promise for biomedical applications, but they are typically manufactured using chemically functionalized surfaces that still require optimization. Here, we describe novel hetero-functionally activated glass surfaces favoring oriented antibody binding for improved performan...

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Bibliographic Details
Published in:Analytical biochemistry 2014-04, Vol.450, p.37-45
Main Authors: González-González, María, Bartolome, Raquel, Jara-Acevedo, Ricardo, Casado-Vela, Juan, Dasilva, Noelia, Matarraz, Sergio, García, Jacinto, Alcazar, J.A., Sayagues, J.M., Orfao, Alberto, Fuentes, Manuel
Format: Article
Language:English
Subjects:
Animals
Antibodies, Immobilized - chemistry
Antibodies, Immobilized - immunology
Antibody arrays
Antibody immobilization
Functionalized surfaces
Glass - chemistry
HLA Antigens - immunology
Humans
Leukocytes, Mononuclear - immunology
Protein Array Analysis - methods
Protein microarrays
Reproducibility of Results
Surface functionalization
Surface Properties
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Summary:Antibody arrays hold great promise for biomedical applications, but they are typically manufactured using chemically functionalized surfaces that still require optimization. Here, we describe novel hetero-functionally activated glass surfaces favoring oriented antibody binding for improved performance in protein microarray applications. Antibody arrays manufactured in our facility using the functionalization chemistries described here proved to be reproducible and stable and also showed good signal intensities. As a proof-of-principle of the glass surface functionalization protocols described in this article, we built antibody-based arrays functionalized with different chemistries that enabled the simultaneous detection of 71 human leukocyte membrane differentiation antigens commonly found in peripheral blood mononuclear cells. Such detection is specific and semi-quantitative and can be performed in a single assay under native conditions. In summary, the protocol described here, based on the use of antibody array technology, enabled the concurrent detection of a set of membrane proteins under native conditions in a specific, selective, and semi-quantitative manner and in a single assay.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2014.01.002