Journal
JOURNAL OF PROTEOME RESEARCH
Volume 5, Issue 7, Pages 1682-1687Publisher
AMER CHEMICAL SOC
DOI: 10.1021/pr0601133
Keywords
mass spectrometry; immunoassay; C-reactive protein; quantitation; multiplexing; human plasma
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Funding
- NCRR NIH HHS [M01-RR000039] Funding Source: Medline
- NIDDK NIH HHS [R18-DK066204, 1 R42DK071290-01] Funding Source: Medline
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Reported in this work is the development and application of a high sensitivity mass spectrometric immunoassay for the quantitative analysis of C-reactive protein from human plasma. Multiplexed affinity retrieval devices and methodology were developed to simultaneously target retinol binding protein, C-reactive protein, serum amyloid P component, as well as an added exogenous internal reference standard (staphylococcal enterotoxin B) for subsequent MALDI-TOF MS analysis. This approach allows for semiquantitative analysis of both retinol binding protein and serum amyloid P component while performing absolute quantitative measurements of C-reactive protein. The ability to qualitatively differentiate between all three human proteins and their associated variants is also maintained. Standard curve, QC, and human plasma samples were analyzed in a high throughput manner, which performed with a CV < 15%. The resultant human plasma sample C-reactive protein quantitative measurements were then compared to those achieved with a high sensitivity latex immunoturbidimetric assay.
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