Production of Monoclonal Antibodies AgainstNeisseria meningitidisUsing Popliteal Lymph Nodes andIn Vivo/In VitroImmunization: Prevalence Study of New Monoclonal Antibodies in Greater São Paulo, Brazil

Abstract

A rapid and efficient method for preparing monoclonal antibody (MAb) serotypes using Neisseria meningitidis outer membrane were used in BALB/c mouse footpads for the immunization. The popliteal lymph nodes were isolated 19 days later for MAb-producing hybridomas, from which the MAbs against the 37 kDa protein were screened. Variations in class 2/3 (PorB) proteins form the basis for meningococcal serotyping. This is the first report on the preparation of MAbs against N. meningitidis that is specific to PorB protein using popliteal lymph nodes. The new monoclonal antibodies were specific for PorB outer membrane protein FL24(PL)Br, a new serotype 24 class 3 antigens of non-typeable (NT:NST) serogroup B strain, and FL14(PL)Br specific for the serotype 14, and reacted with the S3446 reference strain analyzed. A total of 12% of the case isolates reacted with one or more of the monoclonal antibodies. The high-affinity MAbs produced by hybridoma methodology provide a basis for further research on the pathogenesis and early diagnosis of meningococcus.

Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA

“…Here, we describe the use of a large number of Mabs of different isotypes and different specificities for antigens of N. meningitids . Our laboratory has been working for more than 10 years on the development of new monoclonal antibodies against this pathogen [ 9 , 10 , 25 , 35 , 44 ] and also with others pathogens [ 45 - 50 ]. When we need to send our monoclonals to confirm their utility, this becomes an efficient approach.…”

Prime-Boost Immunogenicity of Class 5C Protein from Neisseria meningitidis in Mice with Different Adjuvants

“…The murine monoclonal antibodies (mAb) used for isolation and characterization of the meningococcal variant against Opc were produced in our laboratory as described previously [45] with some modifications [46]. BALB/c female mice (18-22 g -provided by the animal house of Adolfo Lutz Institute) were injected in the footpad with an emulsion of complete Freund's adjuvant containing 5 g of the purified Opc protein.…”

Utilização de anticorpos monoclonais como diferenciais de meningites bacterianas e virais pela técnica de Imuno-histoquímica.

“…Com aplicabilidade na área de Neisseria meningitidis, o sistema de sorotipagem para meningococos baseia-se em uma variedade de anticorpos monoclonais que reconhecem diferenças antigênicas nas proteínas de membrana externa de classe 2 ou 3 (porina B) e de classe 1 (porina A) . O sistema de tipagem com anticorpos monoclonais foram desenvolvidos por causa das dificuldades encontradas com o uso de soros policlonais hiperimunes para tipagem, como descrito em trabalhos publicados pelo nosso laboratório (BELO, 2004(BELO, , 2007CHAVES et al, 2015;DE GASPARI, ZOLLINGER, 2001;FERRAZ et al, 2008). (POOLMAN et al, 1994).…”

“…meningitidis com o painel de anticorpos monoclonais pré-estabelecido e produção de novos monoclonais para a caracterização de perfis desconhecidos (BELO, 2004(BELO, , 2007DE GASPARI, ZOLLINGER, 2001).…”