Quantification the Recombinant SUMO-psTnI in E.coli by SPE-HPLC
- Tu Zhang, Qiu-Ling Xie, Yong Luo, Cai-Kun Wang
- Corresponding Author
- Tu Zhang
Available Online January 2016.
- https://doi.org/10.2991/bst-16.2016.44How to use a DOI?
- SUMO-psTnI, RP-HPLC, SPE/ HPLC, Quantification method, E.coli.
- A simple and rapid reversed-phase liquid chromatography (RP-HPLC) method has been developed and validated for the quantification of recombinant SUMO-psTnI in E.coli BL21/PET-3C. This method could be used to monitor the target protein during fermentation and purification process. After Sonication in 20mMTris buffer for 18 minutes, the supernatant was purified by solid-phase extraction using Ni-NTA column with His tag and then detected on C18 column (250×4.6 mm i.d.,5 m) at 280 nm. The assay was linear over the concentration range 0.30–3.02mg/mL with the correlation coefficients at 0.9953. Standard addition recovery rates of solid-phase extraction from different sample batch were between 91.5% and 98.7%. Relative standard deviation was better than 3.1% .The result showed that the content of SUMO-psTnI inside the bacteria was calculated to be 10.88mg/g. The procedure was rapid and simple to determine the recombinant SUMO-psTnI in E.coli.
- Open Access
- This is an open access article distributed under the CC BY-NC license.
Cite this article
TY - CONF AU - Tu Zhang AU - Qiu-Ling Xie AU - Yong Luo AU - Cai-Kun Wang PY - 2016/01 DA - 2016/01 TI - Quantification the Recombinant SUMO-psTnI in E.coli by SPE-HPLC BT - The International Conference on Biological Sciences and Technology PB - Atlantis Press SN - 2468-5747 UR - https://doi.org/10.2991/bst-16.2016.44 DO - https://doi.org/10.2991/bst-16.2016.44 ID - Zhang2016/01 ER -