A comparative study of CE-SDS, SDS-PAGE, and Simple Western : Influences of sample preparation on molecular weight determination of proteins
© 2020 The Authors. Electrophoresis published by Wiley-VCH GmbH..
The development of capillary electrophoresis, especially CE-SDS devices, has led CE-SDS to become an established tool in a wide range of applications in the analysis of biopharmaceuticals and is increasingly replacing its method of origin, SDS-PAGE. The goal of this study was to evaluate the comparability of molecular weight (MW) determination especially by CE-SDS and SDS-PAGE. For ensuring comparability, model proteins that have little or no posttranslational modifications and an IgG antibody were used. Only a minor influence of sample preparation conditions, including sample buffer, temperature conditions, and different reducing agents on the MW determination were found. In contrast, the selection of the MW marker plays a decisive role in determining the accurate apparent MW of a protein. When using different MW markers, the deviation in MW determination can exceed 10%. Interestingly, CE-SDS and 10% SDS-PAGE hardly differ in their trueness of MW determination. The trueness in relation to the reference MW for each protein was calculated. Although the trueness values for the model proteins considered range between 1.00 and 1.11 using CE-SDS, they range between 0.93 and 1.03 on SDS-PAGE, depending on the experimental conditions chosen.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:42 |
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| Enthalten in: |
Electrophoresis - 42(2021), 3 vom: 04. Feb., Seite 206-218 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Wiesner, Rebecca [VerfasserIn] |
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| Links: |
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| Themen: |
CE-SDS |
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| Anmerkungen: |
Date Completed 18.10.2021 Date Revised 18.10.2021 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1002/elps.202000199 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 245 | 1 | 2 | |a A comparative study of CE-SDS, SDS-PAGE, and Simple Western |b Influences of sample preparation on molecular weight determination of proteins |
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| 500 | |a Date Revised 18.10.2021 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © 2020 The Authors. Electrophoresis published by Wiley-VCH GmbH. | ||
| 520 | |a The development of capillary electrophoresis, especially CE-SDS devices, has led CE-SDS to become an established tool in a wide range of applications in the analysis of biopharmaceuticals and is increasingly replacing its method of origin, SDS-PAGE. The goal of this study was to evaluate the comparability of molecular weight (MW) determination especially by CE-SDS and SDS-PAGE. For ensuring comparability, model proteins that have little or no posttranslational modifications and an IgG antibody were used. Only a minor influence of sample preparation conditions, including sample buffer, temperature conditions, and different reducing agents on the MW determination were found. In contrast, the selection of the MW marker plays a decisive role in determining the accurate apparent MW of a protein. When using different MW markers, the deviation in MW determination can exceed 10%. Interestingly, CE-SDS and 10% SDS-PAGE hardly differ in their trueness of MW determination. The trueness in relation to the reference MW for each protein was calculated. Although the trueness values for the model proteins considered range between 1.00 and 1.11 using CE-SDS, they range between 0.93 and 1.03 on SDS-PAGE, depending on the experimental conditions chosen | ||
| 650 | 4 | |a Comparative Study | |
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a CE-SDS | |
| 650 | 4 | |a Method comparison | |
| 650 | 4 | |a Molecular weight determination | |
| 650 | 4 | |a SDS-PAGE | |
| 650 | 4 | |a Simple Western | |
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| 700 | 1 | |a Krebs, Finja |e verfasserin |4 aut | |
| 700 | 1 | |a Wätzig, Hermann |e verfasserin |4 aut | |
| 700 | 1 | |a Oltmann-Norden, Imke |e verfasserin |4 aut | |
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