DOI: 10.1002/pmic.200500747. Robust protein quantitation in chromatographic fractions using MALDI‐MS of tryptic peptides

Robust protein quantitation in chromatographic fractions using MALDI‐MS of tryptic peptides

10.1002/pmic.200500747

Crossref journal-article

Wiley

PROTEOMICS (311)

Abstract

AbstractA method is introduced to evaluate protein concentrations using the height sum of all MALDI‐MS peaks that unambiguously match theoretic tryptic peptide masses of the protein sought after. The method uses native chromatographic protein fractionation prior to digestion but does not require any depletion, labeling, derivatization, or preparation of a compound similar to the analyte. All peak heights of tryptic peptides are normalized with the peak height of a unique standard peptide added to the MALDI‐MS samples. The sum of normalized peak heights, Sn, or the normalized mean peak height, Mn, reflects the concentration of the respective protein. For fractions containing various proteins, Sn and Mn can be used to compare concentrations of a protein between different fractions. For fractions with one predominating protein, they can be used to estimate concentration ratios between fractions, or to quantify the fractional protein concentration after calibration with pure protein solutions. Initial native fractionation retains the possibility to apply all conventional analytic procedures. Moreover, it renders the method relatively robust to MS mass accuracy. The method was validated with albumin, transferrin, α1‐antitrypsin, and immunoglobulin G within highly complex chromatographic fractions of pathological and normal sera, which contained the respective intact native protein in dominating as well as minor concentrations. The correlation found between Sn and the protein concentration as determined with ELISA showed that the method can be applied to select markers for distinguishing between normal and pathological serum samples.

Bibliography

Bublitz, R., Kreusch, S., Ditze, G., Schulze, M., Cumme, G. A., Fischer, C., Winter, A., Hoppe, H., & Rhode, H. (2006). Robust protein quantitation in chromatographic fractions using MALDIâMS of tryptic peptides. PROTEOMICS, 6(13), 3909â3917. Portico.

Authors 9

Renate Bublitz (first)
Stefan Kreusch (additional)
Günther Ditze (additional)
Margarete Schulze (additional)
Gerhard A. Cumme (additional)
Christine Fischer (additional)
Adrienne Winter (additional)
Horst Hoppe (additional)
Heidrun Rhode (additional)

References 34 Referenced 10

10.1006/abio.1993.1514
10.1073/pnas.90.11.5011
10.1006/bbrc.1993.2009
10.1002/bms.1200220605
10.1016/0960-9822(93)90195-T
10.1002/rcm.1100
10.1002/rcm.734
10.1002/rcm.773
10.1016/S0378-4347(00)00192-4
10.1074/mcp.M400090-MCP200
10.1074/mcp.M400161-MCP200
{'key': 'e_1_2_1_13_2', 'first-page': '662', 'volume': '26', 'author': 'Mirgorodskaia O. A.', 'year': '2000', 'journal-title': 'Bioorg. Khim.'} / Bioorg. Khim. by Mirgorodskaia O. A. (2000)
10.1021/pr025523f
10.1021/ac00063a041
10.1021/ac00081a008
10.1002/jms.1190300219
10.1002/(SICI)1098-2787(1999)18:6<349::AID-MAS1>3.0.CO;2-H
10.1046/j.1432-1327.2000.01019.x
10.1074/jbc.M409587200
10.1074/mcp.M500331-MCP200
10.1002/bms.1200220908
{'key': 'e_1_2_1_23_2', 'first-page': 'S320', 'volume': '4', 'author': 'Gröpl C.', 'year': '2005', 'journal-title': 'Mol. Cell. Proteomics'} / Mol. Cell. Proteomics by Gröpl C. (2005)
10.1021/ac026468x
10.1074/mcp.M400061-MCP200
10.1021/ac048455k
{'key': 'e_1_2_1_27_2', 'first-page': '167', 'volume': '15', 'author': 'Zhang X.', 'year': '2004', 'journal-title': 'J. Biomol. Tech.'} / J. Biomol. Tech. by Zhang X. (2004)
10.1111/j.1432-1033.1993.tb18234.x
10.1002/pmic.200500142
10.1016/S0003-2697(02)00460-8
Horn A. Kreusch S. Sammler G. Bublitz R. Schwedler S. Patent application files DE 10160975.2 WO 03/049841A1 GB 0415432.4
10.1002/pmic.200400873
10.1097/00008480-200404000-00011
10.1159/000068978
10.1053/j.ajkd.2003.10.002

Dates

Type	When
Created	19 years, 2 months ago (May 31, 2006, 5:18 p.m.)
Deposited	1 year, 9 months ago (Nov. 14, 2023, 11:42 p.m.)
Indexed	1 year, 9 months ago (Nov. 15, 2023, 12:19 a.m.)
Issued	19 years, 1 month ago (June 30, 2006)
Published	19 years, 1 month ago (June 30, 2006)
Published Online	19 years, 1 month ago (June 30, 2006)
Published Print	19 years, 1 month ago (July 1, 2006)

Funders 0

None

BibTeX

@article{Bublitz_2006, title={Robust protein quantitation in chromatographic fractions using MALDI‐MS of tryptic peptides}, volume={6}, ISSN={1615-9861}, url={http://dx.doi.org/10.1002/pmic.200500747}, DOI={10.1002/pmic.200500747}, number={13}, journal={PROTEOMICS}, publisher={Wiley}, author={Bublitz, Renate and Kreusch, Stefan and Ditze, Günther and Schulze, Margarete and Cumme, Gerhard A. and Fischer, Christine and Winter, Adrienne and Hoppe, Horst and Rhode, Heidrun}, year={2006}, month=jun, pages={3909–3917} }

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