RT Journal Article
SR Electronic
T1 A Biotin Label-based Antibody Array for High-content Profiling of Protein Expression
JF Cancer Genomics - Proteomics
JO Cancer Genomics Proteomics
FD International Institute of Anticancer Research
SP 129
OP 141
VO 7
IS 3
A1 RUOCHUN HUANG
A1 WEIDONG JIANG
A1 JEAN YANG
A1 YING QING MAO
A1 YING ZHANG
A1 WEIMING YANG
A1 DONGZI YANG
A1 BRETT BURKHOLDER
A1 RANI FAN HUANG
A1 RUO-PAN HUANG
YR 2010
UL http://cgp.iiarjournals.org/content/7/3/129.abstract
AB Background/Aim: Profiling protein expression on a global scale will have significant impact on biomedical research, particularly in the discovery and development of drugs and biomarkers. Through the years, several antibody array systems have been invented and developed for multiple protein detection. However, a reliable and high-content system for protein profiling from many biological samples has yet been developed. This study aimed to develop a reliable, easy to use and cost effective method to profile protein expression levels in high-content manner with sufficient sensitivity and specificity. Materials and Methods: To address this problem, a high density antibody array was developed and used this technology to uncover the potential biomarkers of ovarian cancer. In this system, biological samples are labeled with biotin. The biotinylated proteins are then incubated with antibody chips. The presence of proteins captured by the antibody chip is detected using streptavidin-conjugated fluorescent dye (Cy3 equivalent) as a reporter. The signals, which are visualized by laser scanning, are normalized using positive, negative, and internal controls. Results: Using this biotin label-based antibody array technology, the expression levels of 507 human, 308 mouse and 90 rat target proteins can be simultaneously detected, including of cytokines, chemokines, adipokines, growth factors, angiogenic factors, proteases, soluble receptors, soluble adhesion molecules, and other proteins in a variety of samples. Most proteins can be detected at pg/ml and ng/ml levels, with a coefficient of variation of less than 20%. Using human biotin-based antibody arrays, we screened the serum expression profiles of 507 proteins in ovarian cancer patients and healthy individuals. A panel of protein expression showed significant difference between normal and cancer samples (p<0.05). By classification analysis and split-point score analysis of these two groups, a small group of proteins were found to be useful in distinguishing ovarian cancer patients from normal subjects. Conclusion: Our results suggest the biotin label-based antibody arrays that we have developed have great potential in applications for biomarker discovery.