1. Biochips that sequentially capture and focus antigens for immunoaffinity MALDI-TOF MS: a new tool for biomarker verification.
- Author
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Brauer HA, Lampe PD, Yasui YY, Hamajima N, and Stolowitz ML
- Subjects
- Acetonitriles, Antigens chemistry, Breast Neoplasms metabolism, Disulfides, Female, Humans, Prealbumin chemistry, Prealbumin isolation & purification, Protein Isoforms, Reproducibility of Results, Antigens isolation & purification, Biomarkers analysis, Immunoassay methods, Protein Array Analysis methods, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods
- Abstract
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.
- Published
- 2010
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