Quantitative multiplexed C-reactive protein mass spectrometric immunoassay |
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Authors: | Kiernan Urban A Addobbati Riccardo Nedelkov Dobrin Nelson Randall W |
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Institution: | Intrinsic Bioprobes, Inc., 625 S. Smith Rd. Ste. 22, Tempe, Arizona 85281, USA. ukiernan@intrinsicbio.com |
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Abstract: | 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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