Profiling of cytokine expression by biotin‐labeled‐based protein arrays

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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.