DOI: 10.1002/pmic.200500142. Multidimensional proteomics of human serum using parallel chromatography of native constituents and microplate technology

Multidimensional proteomics of human serum using parallel chromatography of native constituents and microplate technology

10.1002/pmic.200500142

Crossref journal-article

Wiley

PROTEOMICS (311)

Abstract

AbstractA versatile, multidimensional, and non‐denaturing proteome separation procedure using microplate technology is presented, yielding a digitized image of proteome composition. In the first dimension, the sample under study is separated into 96 fractions by size exclusion chromatography (SEC). In the second dimension, the fractions of the first dimension are transferred by the liquid‐handling device CyBi™‐Well (CyBio AG, Jena, Germany) to 96 parallel anion exchange chromatography columns. In this way the proteins are conserved in their native states and are distributed in 2400 liquid fractions with high recovery rates and sufficient reproducibility. The resulting fractions are subjected to protein quantitation and identification. Spectrophotometrical and immunological methods and enzyme activity measurements are used for quantitation. To identify proteins, the fractions are subjected to MALDI‐MS, and their tryptic digests to both MALDI‐ and LC‐ESI‐MS/MS. All preparation steps except the first are applied in parallel to sets of multiples of 96 samples. The procedure may be refined by adding more separation steps and may be adapted to various protein amounts and to various proteomes. Moreover, the method offers the opportunity to investigate functional protein complexes. The method was applied to separate the normal human serum proteome. Within 255 fractions exhibiting the highest protein concentrations, 742 proteins were identified by LC‐ESI‐MS/MS peptide sequence tags.

Bibliography

Horn, A., Kreusch, S., Bublitz, R., Hoppe, H., Cumme, G. A., Schulze, M., Moore, T., Ditze, G., & Rhode, H. (2006). Multidimensional proteomics of human serum using parallel chromatography of native constituents and microplate technology. PROTEOMICS, 6(2), 559â570. Portico.

Authors 9

Anton Horn (first)
Stefan Kreusch (additional)
Renate Bublitz (additional)
Horst Hoppe (additional)
Gerhard A. Cumme (additional)
Margarete Schulze (additional)
Thomas Moore (additional)
Günter Ditze (additional)
Heidrun Rhode (additional)

References 34 Referenced 22

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10.1016/S0003-2697(02)00460-8
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Dates

Type	When
Created	19 years, 8 months ago (Dec. 9, 2005, 9:46 a.m.)
Deposited	1 year, 11 months ago (Sept. 12, 2023, 8:05 p.m.)
Indexed	11 months, 3 weeks ago (Sept. 3, 2024, 6:37 p.m.)
Issued	19 years, 7 months ago (Jan. 1, 2006)
Published	19 years, 7 months ago (Jan. 1, 2006)
Published Online	19 years, 7 months ago (Jan. 17, 2006)
Published Print	19 years, 7 months ago (Jan. 1, 2006)

Funders 0

None

BibTeX

@article{Horn_2006, title={Multidimensional proteomics of human serum using parallel chromatography of native constituents and microplate technology}, volume={6}, ISSN={1615-9861}, url={http://dx.doi.org/10.1002/pmic.200500142}, DOI={10.1002/pmic.200500142}, number={2}, journal={PROTEOMICS}, publisher={Wiley}, author={Horn, Anton and Kreusch, Stefan and Bublitz, Renate and Hoppe, Horst and Cumme, Gerhard A. and Schulze, Margarete and Moore, Thomas and Ditze, Günter and Rhode, Heidrun}, year={2006}, month=jan, pages={559–570} }

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