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

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 serotyping of outer membrane antigens of N.meningitidis by dot-blot ELISA. METHODS: Monoclonal antibodies...

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Autores principales: Ferraz, Aline S, Belo, Elza FT, Coutinho, Ligia MCC, Oliveira, Ana P, Carmo, Andréia MS, Franco, Daniele L, Ferreira, Tatiane, Yto, André Y, Machado, Marta SF, Scola, Monica CG, De Gaspari, Elizabeth
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2311311/
https://www.ncbi.nlm.nih.gov/pubmed/18325118
http://dx.doi.org/10.1186/1471-2334-8-30
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author Ferraz, Aline S
Belo, Elza FT
Coutinho, Ligia MCC
Oliveira, Ana P
Carmo, Andréia MS
Franco, Daniele L
Ferreira, Tatiane
Yto, André Y
Machado, Marta SF
Scola, Monica CG
De Gaspari, Elizabeth
author_facet Ferraz, Aline S
Belo, Elza FT
Coutinho, Ligia MCC
Oliveira, Ana P
Carmo, Andréia MS
Franco, Daniele L
Ferreira, Tatiane
Yto, André Y
Machado, Marta SF
Scola, Monica CG
De Gaspari, Elizabeth
author_sort Ferraz, Aline S
collection PubMed
description 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 serotyping 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.
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spelling pubmed-23113112008-04-16 Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA Ferraz, Aline S Belo, Elza FT Coutinho, Ligia MCC Oliveira, Ana P Carmo, Andréia MS Franco, Daniele L Ferreira, Tatiane Yto, André Y Machado, Marta SF Scola, Monica CG De Gaspari, Elizabeth BMC Infect Dis Research Article 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 serotyping 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. BioMed Central 2008-03-06 /pmc/articles/PMC2311311/ /pubmed/18325118 http://dx.doi.org/10.1186/1471-2334-8-30 Text en Copyright © 2008 Ferraz et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Ferraz, Aline S
Belo, Elza FT
Coutinho, Ligia MCC
Oliveira, Ana P
Carmo, Andréia MS
Franco, Daniele L
Ferreira, Tatiane
Yto, André Y
Machado, Marta SF
Scola, Monica CG
De Gaspari, Elizabeth
Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA
title Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA
title_full Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA
title_fullStr Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA
title_full_unstemmed Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA
title_short Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA
title_sort storage and stability of igg and igm monoclonal antibodies dried on filter paper and utility in neisseria meningitidis serotyping by dot-blot elisa
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2311311/
https://www.ncbi.nlm.nih.gov/pubmed/18325118
http://dx.doi.org/10.1186/1471-2334-8-30
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