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Biochips that sequentially capture and focus antigens for immunoaffinity MALDI‐TOF MS: A new tool for biomarker verification
Author(s) -
Brauer Heather Ann,
Lampe Paul D.,
Yasui Yutaka Y.,
Hamajima Nobuyuki,
Stolowitz Mark L.
Publication year - 2010
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.201000219
Subject(s) - biochip , linker , chemistry , cleavage (geology) , gene isoform , matrix assisted laser desorption/ionization , chromatography , nanotechnology , combinatorial chemistry , materials science , biochemistry , computer science , organic chemistry , adsorption , desorption , fracture (geology) , composite material , operating system , gene
A novel approach to immunoaffinity MS is described wherein antibodies are appended to a patterned gold Biochip surface. The Biochip surface is patterned with an array of concentric immunocapture zones composed of highly hydrophilic central zones surrounded by moderately hydrophilic zones that reside on a non‐wetting background, with protein attachment via electrochemically cleavable linkers. After linker cleavage, matrix application forms a discrete spot suitable for MALDI‐TOF‐MS. Use of the Biochip to purify transthyretin from human serum allowed a distinct resolution of four disulfide conjugates and one truncated form isoforms with good mass resolution and sensitivity.