Evaluation of surface chemistries for antibody microarrays

• Published in: Analytical biochemistry Vol. 371; no. 1; pp. 105 - 115
• Main Authors: Seurynck-Servoss, Shannon L., White, Amanda M., Baird, Cheryl L., Rodland, Karin D., Zangar, Richard C.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.12.2007
• Subjects: 60 APPLIED LIFE SCIENCES; ANTIBODIES; Antibodies - immunology; Antibody; BASIC BIOLOGICAL SCIENCES; Chemokine CCL5 - immunology; DETECTION; DISEASES; DNA; E-Selectin - immunology; ELISA; ENZYME IMMUNOASSAY; Enzyme-Linked Immunosorbent Assay; EVALUATION; GLASS; Humans; Intercellular Adhesion Molecule-1 - immunology; Male; Microarray; MORPHOLOGY; Oligonucleotide Array Sequence Analysis - instrumentation; Oligonucleotide Array Sequence Analysis - methods; Prostate-Specific Antigen - immunology; PROTEINS; Reproducibility of Results; Sensitivity and Specificity; Solid support; Surface Properties; Solid support; Microarray; Antibody; ELISA
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2007.07.010

Antibody microarrays are an emerging technology that promises to be a powerful tool for the detection of disease biomarkers. The current technology for protein microarrays has been derived primarily from DNA microarrays and is not fully characterized for use with proteins. For example, there are a myriad of surface chemistries that are commercially available for antibody microarrays, but there are no rigorous studies that compare these different surfaces. Therefore, we have used a sandwich enzyme-linked immunosorbent assay (ELISA) microarray platform to analyze 17 different commercially available slide types. Full standard curves were generated for 23 different assays. We found that this approach provides a rigorous and quantitative system for comparing the different slide types based on spot size and morphology, slide noise, spot background, lower limit of detection, and reproducibility. These studies demonstrate that the properties of the slide surface affect the activity of immobilized antibodies and the quality of data produced. Although many slide types produce useful data, glass slides coated with aldehyde silane, poly-l-lysine, or aminosilane (with or without activation with a crosslinker) consistently produce superior results in the sandwich ELISA microarray analyses we performed.