Evaluation of Surface Chemistries for Antibody Microarrays
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 primarily derived 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 no rigorous studies that compare these different surfaces. Therefore, we have used an enzyme-linked immunosorbent assay (ELISA) microarray platform to analyze 16 different commercially available slide types. Full standard curves were generated for 24 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 can produce useful data, glass slides coated with poly-L-lysine or aminosilane, with or without activation with a crosslinker, consistently produce superior results in the ELISA microarray analyses we performed.
- Research Organization:
- Pacific Northwest National Laboratory (PNNL), Richland, WA (US)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 921405
- Report Number(s):
- PNNL-SA-55339; 400412000
- Journal Information:
- Analytical Biochemistry, 371(1):105-115, Journal Name: Analytical Biochemistry, 371(1):105-115 Journal Issue: 1 Vol. 371; ISSN ANBCA2; ISSN 0003-2697
- Country of Publication:
- United States
- Language:
- English
Similar Records
Immobilization strategies for single-chain antibody microarrays
High-Throughput Analysis of Serum Antigens Using Sandwich ELISAs on Microarrays