Quantification the Recombinant SUMO-psTnI in E.coli by SPE-HPLC

DOI

10.2991/bst-16.2016.44How to use a DOI?

Keywords

SUMO-psTnI, RP-HPLC, SPE/ HPLC, Quantification method, E.coli.

Abstract

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.

Copyright

© 2016, the Authors. Published by Atlantis Press.

Open Access

This is an open access article distributed under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/).

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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  - Proceedings of the 2016 International Conference on Biological Sciences and Technology
PB  - Atlantis Press
SP  - 295
EP  - 302
SN  - 2468-5747
UR  - https://doi.org/10.2991/bst-16.2016.44
DO  - 10.2991/bst-16.2016.44
ID  - Zhang2016/01
ER  -