Jing Zhang
Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Xi`an, China
Chenhui Zhu
Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Xi`an, China
Daidi Fan
Shaanxi Key Laboratory of Degradable Biomedical Materials, School of Chemical Engineering, Northwest University, Xi`an, China
ABSTRACT
Endotoxin removal from recombinant proteins is considered to be an indispensable step in bioprocesses before the application in biomaterials for pharmaceutical industry. This study was carried out using nonionic detergent Triton X-114 to eliminate endotoxin molecules from human-like collagen preparations. With the evaluation of four factors (Triton X-114 concentration, temperature, time and pH) which were mainly affecting the two-phase extraction behavior, the optimal experimental condition was determined. Under this condition, nearly 98~99% endotoxins were removed with high protein recovery (95.1%). The residual Triton X-114 was efficiently separated from HLC preparation via ultrafiltration procedure with microgram amounts of residues left. Through toxicity test in vitro, the processed HLC was proved to be non-toxic to BHK21 and acceptable as a novel biomaterial. Therefore, Triton X-114 two-phase extraction can be used as an effective way to remove endotoxins from HLC preparations.
PDF References
How to cite this article
Jing Zhang, Chenhui Zhu and Daidi Fan, 2013. Endotoxin Removal from Recombinant Human-like Collagen Preparations by Triton X-114 Two-phase Extraction. Biotechnology, 12: 135-139.
DOI: 10.3923/biotech.2013.135.139
URL: https://scialert.net/abstract/?doi=biotech.2013.135.139
DOI: 10.3923/biotech.2013.135.139
URL: https://scialert.net/abstract/?doi=biotech.2013.135.139
REFERENCES
- Bito, L.Z., 1977. Inflammatory effects of endotoxin-like contaminants in commonly used protein preparations. Science, 196: 83-85.
PubMedDirect Link - Liu, S., R. Tobias, S. McClure, G. Styba, Q. Shi and G. Jackowski, 1997. Removal of endotoxin from recombinant protein preparations. Clin. Biochem., 30: 455-463.
CrossRef - Lopes, A.M., P.O. Magalhaes, P.G. Mazzola, C.O. Rangel-Yagui, J.C. M. de Carvalho, T.C.V. Penna and A. Pessoa, 2010. LPS removal from an E. coli fermentation broth using aqueous two-phase micellar system. Biotechnol. Prog., 26: 1644-1653.
CrossRef - Petsch, D. and F.B. Anspach, 2000. Endotoxin removal from protein solutions. J. Biotechnol., 76: 97-119.
PubMedDirect Link - Weber, J.R., E.F. Degner and K.S. Bahjat, 1964. Determination of Nonionic ethylene oxide adduct in some commercial products. Anal. Chem., 36: 678-679.
CrossRef - Yang, X.J., C.Y. Liang, Y.L. Cai, K. Hu, Q. Wei and Z.D. Cui, 2009. Recombinant human-like collagen modulated the growth of nano-hydroxyapatite on NiTi alloy. Mater. Sci. Eng.: C, 29: 25-28.
CrossRef - Zhu, C.H., D.D. Fan, X.X. Ma, W.J. Xue and Y.Y. Yu et al., 2009. Effects of effects of chitosan on properties of novel human-like collagen/chitosan hybrid vascular scaffold. J. Bioact. Compat. Polym., 24: 560-576.
CrossRef