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Liquid phase separation of proteins based on electrophoretic effects in an electrospray setup during sample introduction into a gas-phase electrophoretic mobility molecular analyzer (CE-GEMMA/CE-ES-DMA).

Weiss VU, Kerul L, Kallinger P, Szymanski WW, Marchetti-Deschmann M, Allmaier G - Anal. Chim. Acta (2014)

Bottom Line: This makes the EM determination of individual species sometimes difficult, if not impossible.This finding was consecutively applied for on-line desalting allowing EM diameter determination of analytes despite a high salt concentration within samples.Results demonstrate the proof of concept of such an approach and additionally illustrate the high potential of a future on-line coupling of a capillary electrophoresis to a GEMMA instrument.

View Article: PubMed Central - PubMed

Affiliation: Institute of Chemical Technologies and Analytics, Vienna University of Technology, Vienna, Austria.

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Related in: MedlinePlus

CE separation of BSA and IgG in CM: the sample contained both analytes, BSA and IgG at c = 1 μmol L−1 protein concentration and pH 9.4 ammonium acetate, respectively. Measurement conditions as in Fig. 3, median values from 6 individual measurements are shown. The IgG peak is detected in scan 15, BSA in scan 16. Hence, CE separation of analytes can be demonstrated.
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fig0025: CE separation of BSA and IgG in CM: the sample contained both analytes, BSA and IgG at c = 1 μmol L−1 protein concentration and pH 9.4 ammonium acetate, respectively. Measurement conditions as in Fig. 3, median values from 6 individual measurements are shown. The IgG peak is detected in scan 15, BSA in scan 16. Hence, CE separation of analytes can be demonstrated.

Mentions: In a next step we mixed both proteins in a single sample and tried to resolve the two analytes via electrophoresis in the liquid phase in CM. Fig. 5 shows consecutive single GEMMA scans demonstrating the separation of the two proteins, BSA and IgG, from such a mixed sample. A peak corresponding to IgG with significant height can be found just in scan 15. BSA is detected only in scan 16. Small traces of IgG can be observed in scans 14 and 16, respectively. This can be explained by a broadly distributed IgG peak resulting from analyte molecules retained on the capillary wall or tip. A corresponding blank (data not shown) only gives a baseline signal. Nevertheless, it can be concluded that analytes injected from a single sample to a commercially available, conventional GEMMA instrument can indeed be separated by CE in the nano ES capillary. A true CE–GEMMA hyphenation is given in CM.


Liquid phase separation of proteins based on electrophoretic effects in an electrospray setup during sample introduction into a gas-phase electrophoretic mobility molecular analyzer (CE-GEMMA/CE-ES-DMA).

Weiss VU, Kerul L, Kallinger P, Szymanski WW, Marchetti-Deschmann M, Allmaier G - Anal. Chim. Acta (2014)

CE separation of BSA and IgG in CM: the sample contained both analytes, BSA and IgG at c = 1 μmol L−1 protein concentration and pH 9.4 ammonium acetate, respectively. Measurement conditions as in Fig. 3, median values from 6 individual measurements are shown. The IgG peak is detected in scan 15, BSA in scan 16. Hence, CE separation of analytes can be demonstrated.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150488&req=5

fig0025: CE separation of BSA and IgG in CM: the sample contained both analytes, BSA and IgG at c = 1 μmol L−1 protein concentration and pH 9.4 ammonium acetate, respectively. Measurement conditions as in Fig. 3, median values from 6 individual measurements are shown. The IgG peak is detected in scan 15, BSA in scan 16. Hence, CE separation of analytes can be demonstrated.
Mentions: In a next step we mixed both proteins in a single sample and tried to resolve the two analytes via electrophoresis in the liquid phase in CM. Fig. 5 shows consecutive single GEMMA scans demonstrating the separation of the two proteins, BSA and IgG, from such a mixed sample. A peak corresponding to IgG with significant height can be found just in scan 15. BSA is detected only in scan 16. Small traces of IgG can be observed in scans 14 and 16, respectively. This can be explained by a broadly distributed IgG peak resulting from analyte molecules retained on the capillary wall or tip. A corresponding blank (data not shown) only gives a baseline signal. Nevertheless, it can be concluded that analytes injected from a single sample to a commercially available, conventional GEMMA instrument can indeed be separated by CE in the nano ES capillary. A true CE–GEMMA hyphenation is given in CM.

Bottom Line: This makes the EM determination of individual species sometimes difficult, if not impossible.This finding was consecutively applied for on-line desalting allowing EM diameter determination of analytes despite a high salt concentration within samples.Results demonstrate the proof of concept of such an approach and additionally illustrate the high potential of a future on-line coupling of a capillary electrophoresis to a GEMMA instrument.

View Article: PubMed Central - PubMed

Affiliation: Institute of Chemical Technologies and Analytics, Vienna University of Technology, Vienna, Austria.

Show MeSH
Related in: MedlinePlus