Mass spectrometric immunoassay for quantitative determination of protein biomarker isoforms.
Protein biomarkers are essential in assessing pathogenic processes. The impetus for finding new biomarkers has been accelerated by the arrival of the "omics" technologies. However, equally important is the rediscovery of existing biomarkers with these new approaches as novel variants can be discovered that can improve their utility. Presented here is a mass spectrometric immunoassay method for quantitative determination of β-2-microglobulin, an established biomarker used in the diagnosis of active rheumatoid arthritis and kidney disease, and its structural variant, cleaved at and deficient in lysine-58 (ΔK58-b2m). β-Lactoglobulin was incorporated into the assay as an internal reference standard, serving as normalization point for β-2-microglobulin quantification. The precision, linearity, and recovery characteristics of the assay were established. The new assay was also benchmarked against existing β-2-microglobulin ELISA. The assay was utilized to determine the individual concentration of β-2-microglobulin and its variant across a larger cohort of samples, demonstrating the ability to simultaneously quantify both proteins.
Trenchevska
Olgica
O
Intrinsic Bioprobes Inc., Tempe, AZ 85284, USA.
Kamcheva
Elena
E
Nedelkov
Dobrin
D
D001172:Arthritis, Rheumatoid / Q000175:diagnosis; D015415:Biomarkers / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007118:Immunoassay / Q000379:methods* / Q000592:standards; D007674:Kidney Diseases / Q000175:diagnosis; D013058:Mass Spectrometry / Q000379:methods* / Q000592:standards; D020033:Protein Isoforms / Q000032:analysis*; D012015:Reference Standards; D015203:Reproducibility of Results; D001613:beta 2-Microglobulin / Q000032:analysis / Q000592:standards
D015415:Biomarkers; D020033:Protein Isoforms; D001613:beta 2-Microglobulin
Circ Heart Fail. 2008 Nov;1(4):258-64
19808300
Amyloid. 2009;16(3):151-73
19657763
J Exp Med. 1973 Dec 1;138(6):1608-12
4128442
Clin Chem. 2010 Feb;56(2):177-85
19884488
Anal Biochem. 2001 Feb 1;289(1):26-35
11161291
Clin Chim Acta. 1984 Aug 15;141(1):41-50
6088125
Ann Rheum Dis. 1978 Aug;37(4):328-32
80161
Nat Methods. 2009 Jun;6(6):411-2
19448640
Eur J Biochem. 1990 Apr 30;189(2):423-9
2110898
J Urol. 2007 Jul;178(1):10-1
17574050
Eur J Biochem. 1987 Feb 16;163(1):21-8
3028795
FEBS J. 2006 Jun;273(11):2461-74
16704420
Brief Funct Genomic Proteomic. 2008 Sep;7(5):355-70
19106161
Biochemistry. 2005 Mar 22;44(11):4397-407
15766269
Proc Natl Acad Sci U S A. 2005 Aug 2;102(31):10852-7
16043703
Nat Biotechnol. 2009 Jul;27(7):633-41
19561596
Clin Chem. 2008 Nov;54(11):1749-52
18957555
Scand J Immunol. 1979 Mar;9(3):247-54
86201
Kidney Int. 1987 Nov;32(5):635-41
3323598
Anal Chem. 2001 Jul 15;73(14):3294-9
11476228
J Diabetes Sci Technol. 2009 Jul 01;3(4):629-34
20144304
Mol Cell Proteomics. 2007 Jul;6(7):1183-7
17468123
Scand J Immunol. 2009 Mar;69(3):203-12
19281532