دورية أكاديمية
Comparison of protein precipitation methods for sample preparation prior to proteomic analysis of Chinese hamster ovary cell homogenates
| العنوان: | Comparison of protein precipitation methods for sample preparation prior to proteomic analysis of Chinese hamster ovary cell homogenates |
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| المؤلفون: | Saumel Pérez-Rodriguez, Octavio T. Ramírez, Mauricio A. Trujillo-Roldán, Norma A. Valdez-Cruz |
| المصدر: | Electronic Journal of Biotechnology, Vol 48, Iss , Pp 86-94 (2020) |
| بيانات النشر: | Elsevier, 2020. |
| سنة النشر: | 2020 |
| المجموعة: | LCC:Biotechnology LCC:Biology (General) |
| مصطلحات موضوعية: | Cell homogenates, Chinese hamster ovary cells, Precipitation protocols, Protein precipitation, Protein recovery, Protein solubilization, Biotechnology, TP248.13-248.65, Biology (General), QH301-705.5 |
| الوصف: | Background: Chinese hamster ovary (CHO) cells are the workhorse for obtaining recombinant proteins. Proteomic studies of these cells intend to understand cell biology and obtain more productive and robust cell lines for therapeutic protein production in the pharmaceutical industry. Because of the great importance of precipitation methods for the processing of samples in proteomics, the acetone, methanol-chloroform (M/C), and trichloroacetic acid (TCA)-acetone protocols were compared for CHO cells in terms of protein recovery, band pattern resolution, and presence on SDS-PAGE. Results: Higher recovery and similar band profile with cellular homogenates were obtained using acetone precipitation with ultrasonic bath cycles (104.18 ± 2.67%) or NaOH addition (103.12 ± 5.74%), compared to the other two protocols tested. TCA-acetone precipitates were difficult to solubilize, which negatively influenced recovery percentage (77.91 ± 8.79%) and band presence. M/C with ultrasonic homogenization showed an intermediate recovery between the other two protocols (94.22 ± 4.86%) without affecting protein pattern on SDS-PAGE. These precipitation methods affected the recovery of low MW proteins ( |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 0717-3458 |
| Relation: | http://www.sciencedirect.com/science/article/pii/S0717345820300452; https://doaj.org/toc/0717-3458 |
| DOI: | 10.1016/j.ejbt.2020.09.006 |
| URL الوصول: | https://doaj.org/article/26e5750e988c4249a3220ea6bf1001c0 |
| رقم الأكسشن: | edsdoj.26e5750e988c4249a3220ea6bf1001c0 |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 07173458 |
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| DOI: | 10.1016/j.ejbt.2020.09.006 |