title: Profiling of cytokine expression by biotin-labeled-based protein arrays.
abstract: 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.
authors.affiliation: Department of Gynecology and Obstetrics, Emory University School of Medicine, Atlanta, GA, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Lin
authors.forename: Ruochun
authors.initials: R
authors.lastname: Huang
authors.forename: Li-Pai
authors.initials: LP
authors.lastname: Chen
authors.forename: Henry
authors.initials: H
authors.lastname: Lisoukov
authors.forename: Zhen-Hai
authors.initials: ZH
authors.lastname: Lu
authors.forename: Shiyong
authors.initials: S
authors.lastname: Li
authors.forename: Cheng C
authors.initials: CC
authors.lastname: Wang
authors.forename: Ruo-Pan
authors.initials: RP
authors.lastname: Huang
mesh_terms: 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
chemical_list: D000906:Antibodies; D016207:Cytokines; D011960:Receptors, Estrogen