Cabrera O, Pérez R, Acosta M, Cabrera RA, Álvarez M, Pérez O, Cuello M

Language: Spanish
References: 16
Page: 22-27
PDF size: 523.27 Kb.

ABSTRACT

The search of adjuvants has been one of the most important lines of the research in the development of vaccines in the last decades. One of the adjuvants under research at Finlay Institute uses lipopolysaccharide (LPS) from Neisseria meningitidis serogroup B in its composition and it is very toxic. For this reason we aim to make a structural modification of LPS from N. meningitidis serogroup B to reduce its toxicity and to study its stability for three months. A LPS purified by water and phenol extraction method was modified structurally by a controlled basic hydrolysis. The modified LPS was controlled at moment of the hydrolysis, a month and three months later. The evaluations consisted on the determination the concentrations of amine groups generated, the toxicity by Limulus Amoebocyte Lysate (LAL), the determination of the electrophoretic pattern and the constant of distribution (Kd) by chromatography of molecular exclusion as indicative parameters of structural integrity. The obtained results showed an increase of amine groups, a reduction of more than 5 000 times of the initial toxicity of native LPS and an increase of the Kd determined by chromatography. Studied parameters remained constant in the three studied months. In summary, modified structurally LPS by means of a controlled basic hydrolysis and stored in watery solution and temperatures from 2 to 8 °C remains without changes in their structure at least for three months.

REFERENCES

1. Mueller JEL, Sangare´BM, Njanpop-Lafourcade Z, Tarnagda Y, Traore S, Yaro R, et al. Molecular characteristics and epidemiology of meningococcal carriage, Burkina Faso. Emerg Infect Dis 2003;13:847-54.

2. Rietschel ET, Westphal O. History of Endotoxins. In: Brade H, Opal SM, Vogel SN, Morrison DC, editors. Endotoxins in Health and Disease. New York: Marcel Dekker; 1999. p.1-30.

3. Ormeño-Orrillo E. Lipopolisacáridos de Rhizobiaceae: estructura y biosíntesis. Rev Latinoamericana de Microbiología 2005;47(3-4):165-75.

4. Raetz CRH, Whitfield C. Lipopolysaccharide endotoxins. Annu Rev Biochem 2002;71:635-700.

5. Jaques M. "Role of lipo-oligosaccharides and lipopolysacharides in bacterial adherence". Trends and Microbiology 1996;4:408-10

6. Harvill E, Preston A, Cotter P, Allen A, Maskell D, Miller J. "Multiple roles for Bordetella lipopolysaccharide molecules during respiratory tract infection". Infect and Immun 2000;68:6720-8.

7. Kahler CM, Martín LE, Shih GC, Rahman MM, Carlson RW, Stephens DS. The (2- 8)-Linked Polysialic Acid Capsule and Lipooligosaccharide Structure Both Contribute to the Ability of Serogroup B Neisseria meningitidis to Resist the Bactericidal Activity of Normal Human Serum. Infect and Immun 1998;66(12):5939-47.

8. Niwa M, Milner KC, Ribi E, Rudbach JA. Alteration of physical, chemical, and biological properties of endotoxin by treatment with mild alkali. J Bacteriol 1969;97:1069-74.

9. Balboa JA, Estrada J, Nápoles LD, González H, Hernández D, Aranguren Y, et al. Purificación de lipopolisacárido de Neisseria meningitidis a partir de una fracción colateral del proceso de producción de VA-MENGOC-BC . Vaccimonitor 2008;17(1):17-26.

10. Cabrera O, Martínez ME, Cuello M, Soto CR, Valmaseda T, Cedré B, et al. Preparation and evaluation of Vibrio cholerae O1. EL Tor Ogawa lipopolysaccharide-tetanus toxoid conjugates. Vaccine 2006;24(S2):74-5.

11. Osborn MJ. Studies on the gran-negative cell in the lipopolysaccharides of Salmonella typhimurium. Proc Natl Acad Sci USA 1963;50:499-506.

12. Church FC, Porter DH, Calignani GL, Swaisgood HE. An ophthalaldehyde spectrophotometric assay for proteinases. Anal Biochem 1986;146(2):343-8.

13. Weary M. Pyrogen Testing of parenteral products-status report. J Parenteral Sci Technol 1984;38(1):20-3.

14. Laemmli UK. Cleavage of structural proteins during the assambly of the head of bacteriophage T4. Nature 1970; 227:680-5.

15. British Pharmacopoeia. Volumen V Production and Testing of Vaccines. Appendix XV: Composition of Polysaccharide Vaccines. Ph. Eur. Method 2.5.31. London, UK: British Pharmacopoeia Online (BP) Edition. 2012. Disponible en: http//bp2012.infostar.com.cn/Bp2012.aspx?tab=browse&a= display&n=6&id=874.

16. Cabrera O. Neoglicoconjugados de origen bacteriano con fines vacunales obtenidos por un nuevo método (Tesis Doctoral). La Habana, Cuba: Facultad de Química, Universidad de La Habana; 2005.