Biotechnology and Bioprocess Engineering:BBE

  • 제11권3호
  • /
  • Pages.215-222
  • /
  • 2006
  • /
  • 1226-8372(pISSN)
  • /
  • 1976-3816(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Optimization of the Lowry Method of Protein Precipitation from the H. influenzae Type b Conjugate Vaccine Using Deoxycholic Acid and Hydrochloric Acid

  • 발행 : 2006.06.30
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

The Lowry method was used in this study to measure protein in Haemophilus influenzae type b (Hib) conjugate vaccines (polyribosylibitol phosphate-tetanus toxoid; PRP-TT) using deoxycholic acid (DOC) to induce protein precipitation. Trichloroacetic acid (TCA) did not induce precipitation adequately from the Hib conjugate bulk and the freeze-dried Hib conjugate product. Its yield was approximately 50%. The matrix structure of Hib conjugate inhibits precipitation by TCA. Although the Lowry method can be carried out without precipitation in Hib conjugate bulk when no residual impurities (adipic acid dihydrazide [ADH], 1-ethyl-3-(3-dimethylamino-propyl) carbodiimide-HCI [EDAC], phenol and cyanogens bromide [CNBr], etc.) are present, it cannot be used for Hib conjugate products that contain sucrose 8.5%, because 8.5% concentration of sucrose enhanced the protein concentration. DOC- and HCl-induced precipitation is an alternative method for evaluating the protein content of the Hib conjugate bulk and the Hib conjugate product. The precipitation was optimal with a final concentrate of 0.1% for DOC at $4^{\circ}C$ and pH 2. This Lowry method, using DOC/HCI precipitation to induce protein precipitation, was confirmed a consistent, reproducible, and valid test for proteins in Hib conjugate bulk and its freeze-dried product.

키워드

참고문헌

  1. Dajani, A. S., B. I. Asmar, and M. C. Thirumoorthi (1979) Systemic Haemophilus influenzae disease: an overview. J. Pediatr. 94: 355-364 https://doi.org/10.1016/S0022-3476(79)80571-5
  2. Granoff, D. M. and M. Basden (1980) Haemophilus influenzae infections in Fresno County, California: a prospective study of the effects of age, race, and contact with a case on incidence of disease. J. Infect. Dis. 141: 40-46 https://doi.org/10.1093/infdis/141.1.40
  3. Todd, J. K. and F. W. Bruhn (1975) Severe Haemophilus influenzae infections. Am. J. Dis. Child. 129: 607-611 https://doi.org/10.1001/archpedi.1975.02120420047016
  4. Cochi, S. L., C. V. Broome, and A. W. Hightower (1985) Immunization of US children with Haemophilus influenzae type b polysaccharide vaccine. A cost-effectiveness model of strategy assessment. J. Am. Med. Ass. 253: 521-529 https://doi.org/10.1001/jama.253.4.521
  5. Coulehan, J. L., R. H. Michaels, K. E. Williams, D. K. Lemley, C. Q. North, Jr., T. K. Welty, and K. D. Rogers (1976) Bacterial meningitis in Navajo Indians. Public Health Rep. 91: 464-468
  6. Tarr, P. I. and G. Peter (1978) Demographic factors in the epidemiology of Haemophilus influenzae meningitis in young children. J. Pediatr. 92: 884-888 https://doi.org/10.1016/S0022-3476(78)80353-9
  7. Ward, J. (1991) Prevention of invasive Haemophilus influenzae type b disease: lessons from vaccine efficacy trials. Vaccine 9 Suppl: S17-S24 https://doi.org/10.1016/0264-410X(91)90175-6
  8. Pittman, M. (1933) The action of type-specific Haemophilus influenzae antiserum. J. Exp. Med. 58: 683-706 https://doi.org/10.1084/jem.58.6.683
  9. Onodera, M. (2002) Silicone rubber membrane bioreactors for bacterial cellulose production. Biotechnol. Bioprocess Eng. 7: 289-294 https://doi.org/10.1007/BF02932838
  10. Barrett, D. J. (1985) Human immune responses to polysaccharide antigens: an analysis of bacterial polysaccharide vaccines in infants. Adv. Pediatr. 32: 139-158
  11. Landsteinder, K. (1962) The Specificity of Serologic Reactions. Cambridge, Harvard University Press, Dover Publications, New York, NY, USA
  12. Bae C. S., S. S. Hong, S. J. Ahn, Y. S. Jang, and B. K. Hur (2005) Importance of the degree of antigen polymerization by detoxification in modulating the immunogenicity of acellular pertussis vaccine. Biotechnol. Bioprocess Eng. 10: 230-235 https://doi.org/10.1007/BF02932018
  13. Dick, W. E. and J. M. Beurret (1989) Glycoconjugates of bacterial carbohydrate antigens: A survey and consideration of design and preparation factors. pp. 48-113. In: J. M. Cruse and R. E. Lewis (eds.). Conjugate Vaccines Contrib. Microb. Immunol. Karger, Switzerland
  14. Lovrien, R. and D. Matulis (2003) Assays for total protein. pp. 18-24. In: J. E. Coligan, B. M. Duun, D. W. Speicher, and P. T. Wingfield (eds.). Short Protocols in Protein Science. John Wiley & Sons, Inc. Canada
  15. Korea Minimum Requirements of Biological Products (1999) Haemophilus influenzae Type b-Tetanus Toxoid Conjugate Vaccine. pp. 181-190. Korea Food & Drug Administration (KFDA), Seoul, Korea
  16. Smith, P. K., R. I. Krohn, G. T. Hermanson, A. K. Mallia, F. H. Gartner, M. D. Provenzano, E. K. Fujimoto, N. M. Goeke, B. J. Olson, and D. C. Klenk (1985) Measurement of protein using bicinchoninic acid. Anal. Biochem. 150: 76-85 https://doi.org/10.1016/0003-2697(85)90442-7
  17. Lowry, O. H., N. J. Rosebrough, A. L. Farr, and R. J. Randall (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193: 265-275
  18. Peterson, G. L. (1983) Determination of total protein. Meth. Enzymol. 91: 95-121 https://doi.org/10.1016/S0076-6879(83)91014-5
  19. Hartree, E. F. (1972) Determination of protein: A modification of the Lowry method that gives a linear photometric response. Anal. Biochem. 48: 422-427 https://doi.org/10.1016/0003-2697(72)90094-2
  20. Peterson, G. L. (1979) Review of the Folin phenol protein quantitation method of Lowry, Rosebrough, Farr, and Randall. Anal. Biochem. 100: 201-220 https://doi.org/10.1016/0003-2697(79)90222-7
  21. Guo, Y.-Y., R. Anderson, J. McIver, R. K. Gupta, and G. R. Siber (1998) A simple and rapid method for measuring unconjugated capsular polysaccharide (PRP) of Haemophilus influenzae type b in PRP-tetanus toxoid conjugate vaccine. Biologicals 26: 33-38 https://doi.org/10.1006/biol.1997.0121
  22. Lei, Q. P., D. H. Lamb, R. Heller, and P. Pietrobon (2000) Quantitation of low level unconjugated polysaccharide tetanus toxoid-conjugate vaccine by HPAEC/PAD following rapid seperation by deoxycholate/HCl. J. Pharm. Biomed. Anal. 21: 1087-1091 https://doi.org/10.1016/S0731-7085(99)00183-1
  23. Tsai, C.-M., X.-X. Gu, and R. A. Byrd (1994) Quantification of polysaccharide in Haemophilus influenzae type b conjugate and polysaccharide vaccines by high-performance anion-exchange chromatography with pulsed amperometric detection. Vaccine 12: 700-706 https://doi.org/10.1016/0264-410X(94)90219-4
  24. Guidance for Industry, Q2B Validation of Analytical Procedure: Methodology, U.S. Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), Center for Biologics Evaluation and Research (CBER), November 1996, ICH
  25. Guidance for Industry, Biological Method Validation, U.S. Department of Health and Human Services, Food and Drug Administration, Center for Drug Evaluation and Research (CDER), Center for Veterinary Medicine (CVM), May 2001, BP
  26. Annand, R. and P. L. Romeo (1976) Protein in food stuffs. Am. Lab. 8: 37-46
  27. Beyer, R. E. (1983) A rapid biuret assay for protein of whole fatty tissues. Anal. Biochem. 129: 483-485 https://doi.org/10.1016/0003-2697(83)90580-8
  28. Markwell, M. A., S. M. Haas, N. E. Tolbert, and L. L. Bieber (1981) Protein determination in membrane and lipoprotein samples; manual and automated procedures. Meth. Enzymol. 72: 296-303 https://doi.org/10.1016/S0076-6879(81)72018-4
  29. Kim, H. S., T. H. Yoo, Y. S. Jang, H. Kim, J. Y. Park, B. K. Hur, Y. W. Ryu, and J. S. Kim (2004) An animal model to evaluate the protective efficacy of Haemophilus influenzae type b conjugate vaccines. Biotechnol. Bioprocess Eng. 9: 490-494 https://doi.org/10.1007/BF02933491
  30. Yoo, T. H., H. S. Kim, S. S. Park, E. Y. Bang, Y. K. Oh, L. S. Kim, H. Kim, B. K. Hur, Y. W. Ryu, and J. S. Kim (2003) Measurement of free polysaccharide in tetanus toxoid-conjugate vaccine using antibody/ammonium sulfate precipitation. J. Microbiol. Biotechnol. 13: 469-472