Reproducibility of tryptic digestion investigated by quantitative fourier transform ion cyclotron resonance mass spectrometry
(2005) In Journal of Proteome Research 4(2). p.394-399- Abstract
- In this study, the reproducibility of tryptic digestion of complex solutions was investigated using liquid chromatography Fourier transform ion cyclotron resonance (LC FT-ICR) mass spectrometry. Tryptic peptides, from human cerebrospinal fluid, (CSF) were labeled with Quantification-Using-Enhanced-Signal-Tags (QUEST)-markers, or 1-([H-4]nicotinoyloxy)- and 1-([D-4]nicotinoyloxy)-succinimide ester markers. The analysis was performed on abundant proteins with respect-to-intensity ratios and sequence coverage and obtained by comparing differently labeled components from one or different pools. To interpret the dynamics in the proteome, one must be able to estimate the error introduced in each experimental steps. The intra sample variation due... (More)
- In this study, the reproducibility of tryptic digestion of complex solutions was investigated using liquid chromatography Fourier transform ion cyclotron resonance (LC FT-ICR) mass spectrometry. Tryptic peptides, from human cerebrospinal fluid, (CSF) were labeled with Quantification-Using-Enhanced-Signal-Tags (QUEST)-markers, or 1-([H-4]nicotinoyloxy)- and 1-([D-4]nicotinoyloxy)-succinimide ester markers. The analysis was performed on abundant proteins with respect-to-intensity ratios and sequence coverage and obtained by comparing differently labeled components from one or different pools. To interpret the dynamics in the proteome, one must be able to estimate the error introduced in each experimental steps. The intra sample variation due to derivatization was approximately 10%. The inter sample variation depending on derivatization and tryptic digestion was not more than approximately 30%. These experimental observations provide a range for the up- and clown-regulations that are possible to study with electrospray ionization LC FT-ICR mass spectrometry. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/246390
- author
- Hagman, C ; Ramstrom, M ; Bentz, Maria LU ; James, Peter LU ; Hakansson, P and Bergquist, J
- organization
- publishing date
- 2005
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- 1-([D-4]nicotinoyloxy)-succinimide ester markers, 1-([H-4]nicotinoyloxy)- and, quantification, QUEST-markers, LC FT-ICR mass spectrometry, cerebrospinal fluid, tryptic reproducibility
- in
- Journal of Proteome Research
- volume
- 4
- issue
- 2
- pages
- 394 - 399
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- wos:000228421900022
- scopus:17444416139
- ISSN
- 1535-3893
- DOI
- 10.1021/pr049809r
- language
- English
- LU publication?
- yes
- id
- 54a7697c-3395-4b99-99a9-0d24ad803bd7 (old id 246390)
- date added to LUP
- 2016-04-01 12:07:41
- date last changed
- 2023-10-28 06:09:37
@article{54a7697c-3395-4b99-99a9-0d24ad803bd7, abstract = {{In this study, the reproducibility of tryptic digestion of complex solutions was investigated using liquid chromatography Fourier transform ion cyclotron resonance (LC FT-ICR) mass spectrometry. Tryptic peptides, from human cerebrospinal fluid, (CSF) were labeled with Quantification-Using-Enhanced-Signal-Tags (QUEST)-markers, or 1-([H-4]nicotinoyloxy)- and 1-([D-4]nicotinoyloxy)-succinimide ester markers. The analysis was performed on abundant proteins with respect-to-intensity ratios and sequence coverage and obtained by comparing differently labeled components from one or different pools. To interpret the dynamics in the proteome, one must be able to estimate the error introduced in each experimental steps. The intra sample variation due to derivatization was approximately 10%. The inter sample variation depending on derivatization and tryptic digestion was not more than approximately 30%. These experimental observations provide a range for the up- and clown-regulations that are possible to study with electrospray ionization LC FT-ICR mass spectrometry.}}, author = {{Hagman, C and Ramstrom, M and Bentz, Maria and James, Peter and Hakansson, P and Bergquist, J}}, issn = {{1535-3893}}, keywords = {{1-([D-4]nicotinoyloxy)-succinimide ester markers; 1-([H-4]nicotinoyloxy)- and; quantification; QUEST-markers; LC FT-ICR mass spectrometry; cerebrospinal fluid; tryptic reproducibility}}, language = {{eng}}, number = {{2}}, pages = {{394--399}}, publisher = {{The American Chemical Society (ACS)}}, series = {{Journal of Proteome Research}}, title = {{Reproducibility of tryptic digestion investigated by quantitative fourier transform ion cyclotron resonance mass spectrometry}}, url = {{http://dx.doi.org/10.1021/pr049809r}}, doi = {{10.1021/pr049809r}}, volume = {{4}}, year = {{2005}}, }