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Spatially addressable protein array: ssDNA‐directed assembly for antibody microarray
Author(s) -
Ng Jin Kiat,
Ajikumar Parayil Kumaran,
Tang Yew Chung,
Lee Jim Yang,
Stephanopoulos Gregory,
Too HengPhon
Publication year - 2007
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200700183
Subject(s) - protein microarray , lysis , protein array analysis , antigen , antibody microarray , chemistry , dna microarray , microarray , nanotechnology , antibody , combinatorial chemistry , biophysics , materials science , biology , gene , biochemistry , gene expression , genetics , immunology
Protein microarray offers a means for high‐throughput profiling of cellular proteins to provide insights into the mechanisms of biological processes. This study describes the design and fabrication of a robust platform, spatially addressable protein array (SAPA), by exploring the specificity of ssDNA hybridization for self‐assembly of semi‐synthetic ssDNA‐antibody conjugates which capture antigens from complex biological samples. This approach does not involve the direct immobilization of antibodies nor antigen, but instead captures the target antigens in the solution phase followed by self‐directed assembly of the complex onto the surface. In an effort to optimize the platform, the effects of surface chemistry, nonspecific protein adsorption, facile preparation, and purification of ssDNA‐conjugated antibody and capture of the antigen from a complex biological sample such as cell lysate were examined. This platform allowed antigen detection in cell lysate with high sensitivity (1 pM). The method described herein can be extended to the high‐throughput detection of other interacting molecules in solution phase and their subsequent assembly onto any substrate.