Document Detail

Quantitation of Surface-bound Proteins on Biochips Using MALDI-TOF MS.
MedLine Citation:
PMID:  22076340     Owner:  NLM     Status:  In-Data-Review    
We report on a novel method for the quantitation of proteins specifically bound on a ligand-presenting biochip by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). The bound protein was digested by trypsin, and the resulting peptide fragments were analyzed by MALDI-TOF MS in the presence of an isotope-labeled internal standard (IS). The IS has the same sequence as a reference peptide (RP) of the target protein digest, but a different molecular weight. The absolute amount of the specifically bound protein on a biochip is then quantitated by comparison of mass intensities between the RP and the IS. Because they have the same molecular milieu, the mass intensities of these two analytes represent the real amounts of analytes on the chip. As a model system, we tested glutathione s-transferase (GST) and a GST-fusion protein, which were captured on glutathione-presenting biochips. We observed that the glutathione densities on biochips showed a good correlation with the absolute quantity of the proteins. We believe that our method will provide an alternative to currently existing tools for the absolute quantitation of surface-bound proteins.
Juhee Lee; Soo-Ryoon Ryoo; Sang Kyung Kim; Joong-Hoon Ahn; Dal-Hee Min; Woon-Seok Yeo
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Analytical sciences : the international journal of the Japan Society for Analytical Chemistry     Volume:  27     ISSN:  1348-2246     ISO Abbreviation:  Anal Sci     Publication Date:  2011  
Date Detail:
Created Date:  2011-11-14     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8511078     Medline TA:  Anal Sci     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  1127     Citation Subset:  IM    
Department of Bioscience and Biotechnology, Konkuk University.
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