Profiling of cytokine expression by biotin-labeled-based protein arrays.
Global analysis of protein expression holds great promise in basic research and patient care. Previously we demonstrated that multiple cytokines could be detected simultaneously using an enzyme-linked immunosorbent assay protein array system with high sensitivity and specificity. In this paper, we described a biotin-labeled-based protein array system to detect multiple cytokines simultaneously from biological samples. In this new approach, proteins from a variety of biological sources are labeled with biotin. The biotin-labeled proteins are then incubated with antibody chips. Targeted proteins are captured by the array antibodies spotted on the antibody chips. The presence of targeted proteins is detected using Cy3- or Cy5-conjugated streptavidin and signals are imaged by laser scanner. The system also can be easily adapted to a two-color binding assay, allowing measurement of the levels of proteins in a test sample with respect to a reference sample at the same chip. To demonstrate its potential applications, we applied this technology to profile human cytokines, chemokines, growth factors, angiogenic factors and proteases in estrogen receptor (ER)+ and ER- cells. These results suggest that biotin-labeled-based antibody chip technology can provide a practical and powerful means of profiling hundreds or thousands of proteins for research and clinical purposes.
Lin
Ying
Y
Department of Gynecology and Obstetrics, Emory University School of Medicine, Atlanta, GA, USA.
Huang
Ruochun
R
Chen
Li-Pai
LP
Lisoukov
Henry
H
Lu
Zhen-Hai
ZH
Li
Shiyong
S
Wang
Cheng C
CC
Huang
Ruo-Pan
RP
D000906:Antibodies / Q000737:chemistry; D019921:Biotinylation; D001943:Breast Neoplasms / Q000175:diagnosis; D045744:Cell Line, Tumor; D016207:Cytokines / Q000276:immunology / Q000378:metabolism*; D005260:Female; D006801:Humans; D040081:Protein Array Analysis / Q000379:methods*; D011960:Receptors, Estrogen / Q000502:physiology
D000906:Antibodies; D016207:Cytokines; D011960:Receptors, Estrogen