Abstract
Endotoxin removal is critical when producing therapeutic proteins in bacterial systems. This hydrophobic compound can be removed through chromatography or filtration, but presents unique challenges dependent upon protein composition as well as production scale. Here we present a robust method for endotoxin removal at the pilot production scale using fast protein liquid chromatography and buffers specifically engineered for endotoxin removal.
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References
Bone RC (1991) The pathogenesis of sepsis. Ann Intern Med 115:457–469
Glauser MP, Zanetti G, Baumgartner JD, Cohen J (1991) Septic shock: pathogenesis. Lancet 338:732–736
Venet C, Zeni F, Viallon A, Ross A, Pain P, Gery P, Page D, Vermesch R, Bertrand M, Rancon F, Bertrand JC (2000) Endotoxaemia in patients with severe sepsis or septic shock. Intensive Care Med 26:538–544
Morrison DC, Ulevitch RJ (1978) The effects of bacterial endotoxins on host mediation systems. A review. Am J Pathol 93:526–618
Kotani S, Takada H, Tsujimoto M, Ogawa T, Takahashi I, Ikeda T, Otsuka K, Shimauchi H, Kasai N, Mashimo J et al (1985) Synthetic lipid A with endotoxic and related biological activities comparable to those of a natural lipid A from an Escherichia coli re-mutant. Infect Immun 49:225–237
Hack CE, Aarden LA, Thijs LG (1997) Role of cytokines in sepsis. Adv Immunol 66:101–195
Lukacs NW, Hogaboam C, Campbell E, Kunkel SL (1999) Chemokines: function, regulation and alteration of inflammatory responses. Chem Immunol 72:102–120
United States Pharmacopeia—National Formulary (2010) Vol. USP 34-NF29
Jang H, Kim HS, Moon SC, Lee YR, Yu KY, Lee BK, Youn HZ, Jeong YJ, Kim BS, Lee SH, Kim JS (2009) Effects of protein concentration and detergent on endotoxin reduction by ultrafiltration. BMB Rep 42:462–466
Yamamoto C, Kim ST (1996) Endotoxin rejection by ultrafiltration through high-flux, hollow fiber filters. J Biomed Mater Res 32:467–471
Chen RH, Huang C Jr, Newton BS, Ritter G, Old LJ, Batt CA (2009) Factors affecting endotoxin removal from recombinant therapeutic proteins by anion exchange chromatography. Protein Expr Purif 64:76–81
Kunioka M, Choi HJ (1995) Properties of biodegradable hydrogels prepared by gamma-irradiation of microbial poly(epsilon-lysine) aqueous-solutions. J Appl Polym Sci 58:801–806
Morimoto S, Sakata M, Iwata T, Esaki A, Hirayama C (1995) Preparations and applications of polyethyleneimine-immobilized cellulose fibers for endotoxin removal. Polym J 27:831–839
Lowe AJ, Anderson KA, Bardliving CL, Huang C Jr, Teixeira LM, Damasceno LM, Ritter G, Old LJ, Batt CA (2011) Expression and purification of cGMP grade NY-ESO-1 for clinical trials. Biotechnol Prog 27(2):435–441
Matsumae H, Minobe S, Kindan K, Watanabe T, Sato T, Tosa T (1990) Specific removal of endotoxin from protein solutions by immobilized histidine. Biotechnol Appl Biochem 12:129–140
Liu CL, Nikas YJ, Blankschtein D (1996) Novel bioseparations using two-phase aqueous micellar systems. Biotechnol Bioeng 52:185–192
Nikas Y, Liu CL, Abbott N, Blankschtein D (1992) Experimental and theoretical investigations of protein partitioning in 2-phase aqueous micellar systems. Abstr Pap Am Chem Soc 203:140-IEC
Anspach FB, Spille H, Rinas U (1995) Purification of recombinant human basic fibroblast growth factor: stability of selective sorbents under cleaning in place conditions. J Chromatogr A 711:129–139
Karplus TE, Ulevitch RJ, Wilson CB (1987) A new method for reduction of endotoxin contamination from protein solutions. J Immunol Methods 105:211–220
Damais C, Jupin C, Parant M, Chedid L (1987) Induction of human interleukin-1 production by polymyxin B. J Immunol Methods 101:51–56
Chen RH (2009) Increasing inclusion body extractability and recoverability by altering fermentation conditions in high cell density Escherichia coli cultures. In: Chemical engineering, Cornell University, Ithaca
Burgess RR (2009) Refolding solubilized inclusion body proteins. Methods Enzymol 463:259–282
de Marco A (2011) Molecular and chemical chaperones for improving the yields of soluble recombinant proteins. Methods Mol Biol 705:31–51
Eiberle MK, Jungbauer A (2010) Technical refolding of proteins: do we have freedom to operate? Biotechnol J 5:547–559
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Lowe, A.J., Bardliving, C.L., Batt, C.A. (2012). Methods for Chromatographic Removal of Endotoxin. In: Voynov, V., Caravella, J. (eds) Therapeutic Proteins. Methods in Molecular Biology, vol 899. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-921-1_17
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DOI: https://doi.org/10.1007/978-1-61779-921-1_17
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-61779-921-1
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