medigraphic.com
ISSN 1025-0298 (Electronic)
ISSN 1025-028X (Print)

2022, Number 1

<< Back Next >>

VacciMonitor 2022; 31 (1)

Characterization of polyribosylribitol phosphate by high efficiency size exclusion chromatography with ultraviolet detection

Valdés-Cantero A, Méndez-Hernández Y, Cabrales-Rico A, Wood-Duque M, Hernández-Correa J, Díaz-Montel B

Language: Spanish
References: 17
Page: 25-33
PDF size: 505.63 Kb.


ABSTRACT

Haemophilus influenzae type b is an important human pathogen causing some invasive diseases in children less than five years of age. Glycoconjugate vaccines based on polyribosylribitol phosphate have been licensed against this bacterium. Quimi-Hib® is the first and only vaccine against this pathogen using the chemically synthesized polysaccharide. The Active Pharmaceutical Ingredient is produced by the Center for Genetic Engineering and Biotechnology and is obtained from its conjugation to tetanus toxoid. In the present report a characterization of polyribosylribitol phosphate was performed by high performance molecular exclusion chromatography with ultraviolet detection at 215 nm. Three batches were evaluated in the study and the elution profile was determined on a SuperdexTM 75 10/300 GL Increase column with a purity percentage of 77.42 ± 8.97 and an average molecular weight of 7,381 Da ± 210.93. The main impurity present in polyribosylribitol phosphate was dimethylsulfoxide, the solvent used in the activation reaction with N-hydroxysuccinimidyl ester of β-maleimidopropionic acid. Polyribosylribitol phosphate was purified by filtration using a 2,000 Da cut-off Amicon Ultra-15 to a purity of 99.1 % and conjugated to tetanus toxoid. The yield of the conjugation reaction with the purified polysaccharide was 30.0 % ± 1.77 which shows no significant difference with the control which was 33.7%± 3.57 demonstrating that dimethylsulfoxide does not affect the performance of the conjugation reaction.


REFERENCES

  1. Biere E, Rubin L, Moro P, Cohn A, Clark T, Messonnier N. Prevention and Control of Haemophilus influenzae Type b Disease Recommendations of the Advisory Committee on Immunization Practices (ACIP). Morb Mortal Wkly Rep.2014; 63:1-14. Available at: https://www.cdc.gov/mmwr/pdf/rr/rr6301.pdf.

  2. Essam A, Almehdar H, Redwan E. Hib Vaccines: Past, Present, and Future Perspectives. J Immunol Res. 2016; 2016:7203587. doi: https://10.1155/2016/7203587.

  3. Bencomo V. Patente a la invención por Oligosacáridos derivados de ribose-ribitol-fosfato, métodos para prepararlo, inmunógenos que lo comprenden y vacunas que comprenden dicho inmunógeno. US patent 2,252,045.1999 Junio 12.

  4. Lindberg A. Glycoprotein conjugate vaccines. Vaccine.1999; (17 Suppl 2): S28-36.

  5. Anderson PW, Pichichero ME, Stein EC, Porcelli S, Betts RF, Connuck DM, et al. Effect of oligosaccharide chain length, exposed terminal groups, and hapten loading the antibody response of human adults and infants to vaccines consisting of Haemophilus influenzae type b capsular antigen uniterminally coupled to the diphteria protein CRM197. J Immunol 1989; 142(7):2464-8.

  6. Vartak A, Sucheck S. Recent Advances in Subunit Vaccine Carriers. Vaccines (Basel). 2016;4(2):12. doi: https://10.3390/vaccines4020012.

  7. Pollard AJ; Perrett KP; Beverley PC. Maintaining protection against invasive bacteria with protein-polysaccharide conjugate vaccines. Nat Rev Immunol. 2009; 9(3):213-20.

  8. Ambrón LL, Torres LIE, Carreras AP, Santana BMG, Sardiña MÁG, Aguirre SR, Fuentes AT. Experiencia cubana en inmunización,1962-2016. Rev Panam Salud Pública. 2018;42:e34. doi: https://10.26633/RPSP.2018.34.

  9. Centro para el Control Estatal de los Medicamentos (CECMED). Registro sanitario vacuna QuimiHib(r), Report No.: Código B-03-150-J07. La Habana: CECMED; 2003..

  10. Garay HE. Síntesis de péptidos modificados químicamente con posibles aplicaciones farmacéuticas [Tesis doctoral]. La Habana: Universidad de La Habana; 2012.

  11. Soto C, Cuello M, Alfonso Y, Cabrera O, Sierra G. Validación de una técnica colorimétrica para la determinación de carbohidratos. VacciMonitor.2002;11(3):11-4. Disponible en: http://scielo.sld.cu/pdf/vac/v11n3/vac03302.pdf.

  12. Guillén A, Hernández AV, Pérez V, Miranda MP, Pérez NO, López CA. Validación de un método espectrofotométrico para cuantificación de tioles libres en filgrastim. Rev Mex Ing Quim. 2016;15 (3):741-8. doi:https://www.redalyc.org/pdf/620/62048168006.pdf.

  13. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem. 1951; 193:265-75.

  14. Walker JM. The protein Protocols Handbook. Totowa, NJ: Humana Press Inc;2009.

  15. López-Legarda X, Taramuel-Gallardo A, Arboleda Echavarría C, Segura-Sánchez F, Rastrepo-Betancur LF. Comparación de métodos que utilizan ácido sulfúrico para la determinación de azúcares totales. Rev Cub Quim; 2017;29(2):180-98. Disponible en: http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S2224-54212017000200002.

  16. Pérez CS, Ortiz PJ. Espectroscopia. La Habana: Félix Varela; 2010.

  17. Albericio F, Annis I, Royo M, Barany G. Preparation and handling of peptides containing methionine and cysteine. In: Chan WC, White PD. Fmoc solid phase peptide synthesis, A Practical Approach. New York: Oxford University Press inc; 2000.p. 77-109.




2020     |     www.medigraphic.com