Search

Neisseria lactamica, a commensal bacterium that is non-pathogenic to humans and is usually found in the upper respiratory tract of children, is closely related to the pathogenic species Neisseria meningitidis. Colonization by Neisseria lactamica can be responsible for the development of natural immunity to meningococcal infection in childhood, when rates of meningococcal carriers are low. These features suggest that N. lactamica components can be key elements in the production of a new vaccine for N. meningitidis. The production of monoclonal antibodies for N. lactamica is an important tool in the selection of new antigens for the preparation of a vaccine for N. meningitidis B. [ABSTRACT FROM AUTHOR]

Background: A simple filter paper method was developed for, the transport and storage of monoclonal antibodies (Mabs) at room temperature or -20°C after spotting on filter paper, for subsequent sero-typing of outer membrane antigens of N.meningitidis by dot-blot ELISA. Methods: Monoclonal antibodies (Mabs) were spotted within a 0.5-1 cm diameter area of Whatman grade 903 paper, which were stored individually at room temperature or at -20°C. These MAbs were stored and analyzed after periods of one week, 4 weeks, 12 months, or 13 years in the case of frozen Mab aliquots, or after 4 weeks at -20°C or at room temperature (RT) in the case of Mabs dried on filter paper strips. Assays were performed in parallel using dot-blot ELISA. In addition to the MAbs specific for serotyping class 1, 2 or 3, we used a larger number of Mabs for polysaccharides, lipooligosaccharides (LOS), class 5 and cross-reactive antigens for native outer membrane of N.meningitidis. The Mabs dried on filter paper were eluted with phosphate-buffered saline (PBS) containing 0.2% gelatin. Results: Mabs of the isotypes IgG and IgM dried on filter papers were not affected by duration of storage. The detection by serotyping Mabs was generally consistent for dried filter paper MAb samples stored frozen for over 1 year at -20°C, and although decreased reactive antibody titers were found after storage, this did not interfere with the specificity of the Mabs used after 13 years as dry spots on filter paper. Conclusion: The use of filter paper is an inexpensive and convenient method for collecting, storing, and transporting Mab samples for serotyping studies. In addition, the samples occupy little space and can be readily transported without freezing. The efficiency of using immunoglobulin G (IgG) or M (IgM) eluted was found to be consistent with measurement of IgG or IgM titers in most corresponding, ascites Mabs stored frozen for over 1 year. The application of meningococcal typing methods and designations depend on the question being asked. [ABSTRACT FROM AUTHOR]

The article highlights a study on the preparation of monoclonal antibodies (Mab) against Neisseria (N.) meningitidis B. The native outer membrane complex (NOMC) of N. meningitidis B was used in a mice for immunization. The antibodies produced were FL24(PL)Br, a new serotype 24 class 3 antigen of nontypeable serogroup B strain and FL14(PL)Br, which was specific for the serotype 14 and reacted with the S3446 reference strain of N. meningitidis.

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. [ABSTRACT FROM AUTHOR]