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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Effects of protein concentration and detergent on endotoxin reduction by ultrafiltration

  • Jang, Hyun (R&D Center, Komipharm Co.) ;
  • Kim, Hyo-Seung (R&D Center, Komipharm Co.) ;
  • Moon, Seung-Cheol (R&D Center, Komipharm Co.) ;
  • Lee, Young-Rae (Department of Biochemistry and Institute for Medical Sciences, Chonbuk National University Medical School) ;
  • Yu, Kang-Yeoul (Department of Biochemistry and Institute for Medical Sciences, Chonbuk National University Medical School) ;
  • Lee, Byeong-Kil (Department of Surgery, Chonbuk National University Medical School) ;
  • Youn, Hyun-Zo (Department of Surgery, Chonbuk National University Medical School) ;
  • Jeong, Young-Ju (Department of Obstetrics & Gynecology, Chonbuk National University Medical School) ;
  • Kim, Byeong-Soo (Department of Companion and Laboratory Animal Science, Kongju National University College of Medicine) ;
  • Lee, Sung-Ho (Department of Companion and Laboratory Animal Science, Kongju National University College of Medicine) ;
  • Kim, Jong-Suk (Department of Biochemistry and Institute for Medical Sciences, Chonbuk National University Medical School)
  • Published : 2009.07.31
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Abstract

Lipopolysaccharide (LPS), found in the outer membrane of Gram negative bacteria, only exerts its toxic effects when in free form. LPS has three major parts, lipid A, the toxic component, along with a core polysaccharide and O-specific polysaccharide. LPS monomers are known to have molecular masses between 10 to 30 kDa. Under physiological conditions, LPS exists in equilibrium between monomer and vesicle forms. LPS removal by 100 kDa ultrafiltration was more efficient (99.6% of LPS removed) with a low concentration of protein (2.0 mg/ml) compared to a high concentration (20.1 mg/ml). In the presence of different detergents (0.5% Tween 20, 1.0% taurodeoxycholate and 1.0% Triton X-100), LPS removal was more efficient at low protein concentrations (2.0 mg/ml) compared to high protein concentrations (20.1 mg/ml).

Keywords

References

  1. Galanos, C. and Freudenberg, M. A. (1993) Mechanisms of endotoxin shock and endotoxin hypersensitivity. Immunobiology 187, 346-356 https://doi.org/10.1016/S0171-2985(11)80349-9
  2. Raetz, C. R. (1990) Biochemistry of endotoxins. Annu. Rev. Biochem. 59, 129-170 https://doi.org/10.1146/annurev.bi.59.070190.001021
  3. Hannecart-Pokorni, E., Dekegel, D. and Depuydt, F. (1973) Macromolecular structure of lipopolysaccharides from gram-negative bacteria. Eur. J. Biochem. 38, 6-13 https://doi.org/10.1111/j.1432-1033.1973.tb03025.x
  4. Shands, J. W. Jr., Graham, J. A. and Nath, K. (1967) The morphologic structure of isolated bacterial lipopolysaccharide. J. Mol. Biol. 25, 15-21 https://doi.org/10.1016/0022-2836(67)90275-6
  5. Rietschel, E. T., Kirikae, T., Schade, F. U., Mamat, U., Schmidt, G., Loppnow, H., Ulmer, A. J., Zahringer, U., Seydel, U. and Di Padova, F. (1994) Bacterial endotoxin: molecular relationships of structure to activity and function. FASEB. J. 8, 217-225
  6. Baggerman, C., Pathmamanoharan, C., Spies, F., Joosten, J. G. and Junginger, H. E. (1985) Physicochemical properties of endotoxins in large volume parenterals. J. Pharm. Pharmacol. 37, 521-527 https://doi.org/10.1111/j.2042-7158.1985.tb03059.x
  7. Sweadner, K. J., Forte, M. and Nelsen, L. L. (1977) Filtration removal of endotoxin (pyrogens) in solution in different states of aggregation. Appl. Environ. Microbiol. 34, 382- 385
  8. Elass-Rochard, E., Roseanu, A., Legrand, D., Trif, M., Salmon, V., Motas, C., Montreuil, J. and Spik, G. (1995) Lactoferrin-lipopolysaccharide interaction: involvement of the 28-34 loop region of human lactoferrin in the high-affinity binding to Escherichia coli 055B5 lipopolysaccharide. Biochem. J. 312 (Pt 3), 839-845
  9. Koyama, K., Ohno, S. and Murayama, K. (1983) Removal of endotoxin from protein solutions by means of ultrafiltration. Hakko. Kogaku. Kaishi. 61, 421-423
  10. Liu, S., Tobias, R., McClure, S., Styba, G.Shi, Q. and Jackowski, G. (1997) Removal of endotoxin from recombinant protein preparations. Clin. Biochem. 30, 455-463 https://doi.org/10.1016/S0009-9120(97)00049-0
  11. Li, L. and Luo, R. G. (1998) Use of $Ca^{2+}$ to re-aggregate lipopolysaccharide (LPS) in hemoglobin solutions and the subsequent removal of endotoxin by ultrafiltration. Biotechnol. Tech. 12, 119-122 https://doi.org/10.1023/A:1008884332586
  12. Nelsen, L. L. (1978) Removal of Pyrogens from Parenteral Solutions by Ultrafiltration. Pharm. Tech. 2, 46-49
  13. Kastowsky, M., Gutberlet, T. and Bradaczek, H. (1992) Molecular modelling of the three-dimensional structure and conformational flexibility of bacterial lipopolysaccharide. J. Bacteriol. 174, 4798-4806

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