SciELO - Scientific Electronic Library Online

 
vol.9 issue3Guide for the standardization of enzyme immunoassays for use in vaccine trialsDevelopment of Biomodels for the evaluation of the secretory immunity against M tuberculosis in Balb/c mice author indexsubject indexarticles search
Home Pagealphabetic serial listing  

Services on Demand

Journal

Article

Indicators

  • Have no cited articlesCited by SciELO

Related links

  • Have no similar articlesSimilars in SciELO

Share


Vaccimonitor

Print version ISSN 1025-028XOn-line version ISSN 1025-0298

Abstract

ESTRADA, Eric et al. Restoration in Immunoblotting of Neisseria meningitidis proteins denatured by reducing agents and heat. Vaccimonitor [online]. 2000, vol.9, n.3, pp.19-23. ISSN 1025-028X.

Five different detergents for restoration of IgG antibody binding were studied in the washing steps and in the buffer solution used for the dilution of the conjugate and samples. Binding of human serum IgG antibodies to outer membrane protein (OMP) was detected with class-specific peroxidase-conjugated human antibodies. The positive control reacted with those proteins whose molecular weight corresponded approximately to P1, P3, P4 and P5. Proteins of 80 kDa, 70 kDa and 24 kDa and another protein of >150 kDa were also recognized. Our studies indicated that Tween 20 was the best detergent for restoration of OMP antigenicity, except for the 150 kDa protein. Tween 20 achieved a greater number and intensity of bands and a lower background with respect to Empigen BB, Triton X-100, Nonidet NP-40 and CHAPS. The use of detergents affected the antigenicity of the 150 kDa protein. Washing with Tween 20 were the most important steps for restoration of IgG antibody binding sites of heat-denatured OMPs.

Keywords : Immunoblotting; Neisseria meningitidis; detergents; outer membrane proteins; renaturation; denatured proteins.

        · abstract in Spanish     · text in Spanish     · Spanish ( pdf )

 

Creative Commons License All the contents of this journal, except where otherwise noted, is licensed under a Creative Commons Attribution License