Biochips that sequentially capture and focus antigens for immunoaffinity MALDI-TOF MS: a new tool for biomarker verification.
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.
Molecular and Cellular Biology Program, University of Washington, Seattle, WA, USA.
Heather Ann
HA
Brauer
Paul D
PD
Lampe
Yutaka Y
YY
Yasui
Nobuyuki
N
Hamajima
Mark L
ML
Stolowitz
D000097:Acetonitriles; D000941:Antigens / Q000737:chemistry / Q000302:isolation & purification*; D015415:Biomarkers / Q000032:analysis*; D001943:Breast Neoplasms / Q000378:metabolism; D004220:Disulfides; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D011228:Prealbumin / Q000737:chemistry / Q000302:isolation & purification; D040081:Protein Array Analysis / Q000379:methods*; D020033:Protein Isoforms; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D000097:Acetonitriles; D000941:Antigens; D015415:Biomarkers; D004220:Disulfides; D011228:Prealbumin; D020033:Protein Isoforms; C032159:acetonitrile