1. Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA
- Author
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André Yoshio Yto, Andreia Moreira dos Santos Carmo, Ana P. Oliveira, Elza F.T. Belo, Ligia M.C.C. Coutinho, Aline S. Ferraz, Monica C G Scola, Daniele De L. Franco, Elizabeth De Gaspari, Marta S.F. Machado, and Tatiane Ferreira
- Subjects
Paper ,medicine.drug_class ,Dot blot ,Enzyme-Linked Immunosorbent Assay ,Neisseria meningitidis ,Monoclonal antibody ,Immunoglobulin G ,Microbiology ,lcsh:Infectious and parasitic diseases ,Specimen Handling ,Antigen ,medicine ,Animals ,Humans ,lcsh:RC109-216 ,Serotyping ,Chromatography ,biology ,Filter paper ,Micropore Filters ,Antibody titer ,Antibodies, Monoclonal ,Bacterial Typing Techniques ,Infectious Diseases ,Immunoglobulin M ,biology.protein ,Antibody ,Filtration ,Research Article - Abstract
BackgroundA 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 serotyping of outer membrane antigens ofN.meningitidisby dot-blot ELISA.MethodsMonoclonal 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 ofN.meningitidis. The Mabs dried on filter paper were eluted with phosphate-buffered saline (PBS) containing 0.2% gelatin.ResultsMabs 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.ConclusionThe 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.
- Published
- 2007