Quantitative multiplexed C-reactive protein mass spectrometric immunoassay.
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.
Kiernan
Urban A
UA
Intrinsic Bioprobes, Inc., 625 S. Smith Rd. Ste. 22, Tempe, Arizona 85281, USA. ukiernan@intrinsicbio.com
Addobbati
Riccardo
R
Nedelkov
Dobrin
D
Nelson
Randall W
RW
D002097:C-Reactive Protein / Q000032:analysis* / Q000235:genetics; D016022:Case-Control Studies; D003924:Diabetes Mellitus, Type 2 / Q000097:blood; D014644:Genetic Variation; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D011236:Prediabetic State / Q000097:blood; D012015:Reference Standards; D012177:Retinol-Binding Proteins / Q000032:analysis / Q000235:genetics; D054839:Retinol-Binding Proteins, Plasma; D000683:Serum Amyloid P-Component / Q000032:analysis / Q000235:genetics
D012177:Retinol-Binding Proteins; D054839:Retinol-Binding Proteins, Plasma; D000683:Serum Amyloid P-Component; D002097:C-Reactive Protein