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Biochips that sequentially capture and focus antigens for immunoaffinity MALDI‐TOF MS: A new tool for biomarker verification

2010; Wiley; Volume: 10; Issue: 21 Linguagem: Inglês

10.1002/pmic.201000219

1615-9861

Heather Ann Brauer, Paul D. Lampe, Yutaka Yasui, Nobuyuki Hamajima, Mark L. Stolowitz,

Advanced Biosensing Techniques and Applications

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.