Exploring the Complementary Selectivity of Immunocapture and MS Detection for the Differentiation between hCG Isoforms in Clinically Relevant Samples

Whereas numerous immunoassays have been developed to ensure detection of the entire spectrum of isoforms displayed by the human chorionic gonadotropin (hCG) molecule, significant variation has been demonstrated in how these isoforms are recognized by the antibodies in different immunoassays. The aim...

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Bibliographic Details
Published in:Journal of proteome research 2009-11, Vol.8 (11), p.5241-5252
Main Authors: Lund, Hanne, Torsetnes, Silje Bøen, Paus, Elisabeth, Nustad, Kjell, Reubsaet, Léon, Halvorsen, Trine Grønhaug
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Antibodies, Monoclonal - immunology
Biomarkers - chemistry
Biomarkers - metabolism
Chorionic Gonadotropin - blood
Chorionic Gonadotropin - chemistry
Chorionic Gonadotropin - genetics
Chorionic Gonadotropin - urine
Chromatography, Liquid - methods
Female
Humans
Immunoassay - instrumentation
Immunoassay - methods
Immunoassay - standards
Mass Spectrometry - methods
Molecular Sequence Data
Neoplasms - metabolism
Peptides - chemistry
Peptides - genetics
Pregnancy
Protein Isoforms - blood
Protein Isoforms - chemistry
Protein Isoforms - genetics
Protein Isoforms - urine
Proteomics - methods
Sensitivity and Specificity
Solid Phase Extraction - methods
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Summary:Whereas numerous immunoassays have been developed to ensure detection of the entire spectrum of isoforms displayed by the human chorionic gonadotropin (hCG) molecule, significant variation has been demonstrated in how these isoforms are recognized by the antibodies in different immunoassays. The aim of this study was to establish a method using the dual selectivity of the immunoextraction and the mass spectrometry detection for the differentiation between various hCG isoforms in clinically relevant samples. Immunoextraction of endogenous hCG isoforms using a monoclonal antibody (E27) on a 96-well microtitier plate, followed by in-well tryptic digestion, and SIM monitoring of the selected signature peptides, resulted in the qualitative differentiation between several hCG isoforms in serum or urine. We conclude that the orthogonal selectivity conferred by the combination of immunoaffinity extraction and LC-MS analysis offers valuable complementary information to the conventional immunoassays.
ISSN:1535-3893
1535-3907
DOI:10.1021/pr900580n