RESUMO
Methods for the evaluation of the molecular parameters of B polysaccharide (B-PS) in meningococcal protein-polysaccharide vaccine of group B are proposed. The comparison of two proposed methods, the passive hemagglutination inhibition test and rocket immunoelectrophoresis (RIEP), has shown that the latter method has the highest degree of correlation with the chemical method of the detection of B-PS, which is often hindered by lactose added as a bulking agent. RIEP may be recommended for the standardization and control of the commercial preparations of group B meningococcal vaccine. B-PS contained in the vaccine is known to be in a highly polymeric state: its yield is 66.4 +/- 1.7% to Kd = 0.25 and 85.3 +/- 1.2% to Kd = 0.5. B-PS contained in the prepared vaccine has been found to be highly stable. Outer membrane proteins of meningococci, forming a noncovalent complex with capsular B-PS, seem to stabilize its structure and prevent the depolymerization of molecules.
Assuntos
Proteínas da Membrana Bacteriana Externa/imunologia , Vacinas Bacterianas/química , Neisseria meningitidis/imunologia , Polissacarídeos Bacterianos/imunologia , Vacinas Bacterianas/análise , Vacinas Bacterianas/normas , Cromatografia em Gel/métodos , Cromatografia em Gel/estatística & dados numéricos , Contraimunoeletroforese/métodos , Contraimunoeletroforese/estatística & dados numéricos , Testes de Inibição da Hemaglutinação/métodos , Testes de Inibição da Hemaglutinação/estatística & dados numéricos , Imunoeletroforese/métodos , Imunoeletroforese/estatística & dados numéricosRESUMO
New data concerning the influence of cultivation conditions on the synthesis aimed at producing immunologically active antigens of group B Neisseria meningitides were obtained. The combination of conditions (deficiency in ions of iron, pH of the medium, growth phase of the culture) capable of essentially increasing (9- to 28-fold) the content of iron-dependent protein with a mol. wt. of 75-76 kD, was selected. At the same time the content of other immunologically active proteins with mol. wt. of 92-94 kD, 100-102 kD and 141-144 kD perceptibly increased (1.5 to 2.2-fold). Group B meningococcal vaccine prepared on the basis of native protein-polysaccharide complex with a higher content of the above-mentioned species-specific proteins may be expected to have a wider spectrum of immunological action.