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 d...

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Bibliographic Details
Published in:Proteomics (Weinheim) 2003-09, Vol.3 (9), p.1750-1757
Main Authors: Lin, Ying, Huang, Ruochun, Chen, Li-Pai, Lisoukov, Henry, Lu, Zhen-Hai, Li, Shiyong, Wang, Cheng C., Huang, Ruo-Pan
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Antibodies - chemistry
Antibody arrays
Biological and medical sciences
Biotin
Biotinylation
Breast Neoplasms - diagnosis
Cell Line, Tumor
Cytokines
Cytokines - immunology
Cytokines - metabolism
Female
Fundamental and applied biological sciences. Psychology
Humans
Miscellaneous
Protein Array Analysis - methods
Protein chip
Proteins
Receptors, Estrogen - physiology
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Summary: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.
ISSN:1615-9853
1615-9861
DOI:10.1002/pmic.200300530