6 results on '"De Gaspari, Elizabeth"'
Search Results
2. MAb 1D41D8 Against 35 kDa Lipoprotein of Mycoplasma penetrans.
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De Gaspari, Elizabeth N.
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MONOCLONAL antibodies , *MYCOPLASMA , *LIPOPROTEINS , *ANTIGENS , *ENZYME-linked immunosorbent assay , *HYBRIDOMAS , *CELL lines - Abstract
The article provides information on the production of the monoclonal antibody (MAb) 1D41D8 that fights 35 kilodaltons lipoprotein of Mycoplasma penetrans. Lipid-associated membrane proteins (LAMPs) were used as antigen for immunization of a mice. It cites the selection of hybridoma cell lines through the enzyme-linked immunosorbent assay (ELISA). It indicates that such monoclonal antibody recognized 35 kilodalton LAMPs.
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- 2007
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3. Immunogenicity of antigens from outer membrane vesicles of Neisseria meningitidis associated with bilayer fragment of dioctadecyldimethylammonium in Swiss adult mice.
- Author
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Rinaldi, Fabiana Mahylowski, Gaspar, Emanuelle Baldo, Brito, Luciana Tendolini, and De Gaspari, Elizabeth
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VESICLES (Cytology) , *NEISSERIA meningitidis , *LABORATORY mice , *ANTIGENS , *IMMUNOGLOBULIN G , *NEISSERIA , *MICE - Abstract
Purpose: In the present study, meningococcal serogroup B outer membrane vesicles (OMVs) were associated with bilayer fragments of a cationic lipid, dioctadecyldimethylammonium (DDA-BF), used as adjuvant, in an antigenic preparation tested in adult female outbred mice. This adjuvant was compared to the traditional adjuvant aluminum hydroxide. Materials and Methods: The potential in generating humoral response was evaluated by enzyme-linked immunosorbent assay (ELISA). Individual serum was collected and immunoglobulin G (IgG), IgG1, IgG2a, and IgG2b were quantified. Analyses were carried out 15 and 60 days after immunization. Antibodies avidity index were also analyzed by ELISA. Immunoblot and dot-ELISA were carried out to evaluate specific reaction for homologous strains and crossreactive antigens present in other meningococcal strains isolated in 2011-2012 year, in Brazil. Delayed type hypersensitivity was used as indicative of cellular immunity and compared between two experimental groups, 24 hours after homologous strain challenge. Results: The OMVs of Neisseria meningitidis, and N. lactamica (related species) were characterized by electrophoretic separation of proteins in 13% polyacrylamide gel. The strains presented antigens in the range of 8 to 130 kDa, showing a heterogeneous protein migration pattern. In the group immunized with OMVs/DDA-BF, we found no significant production of total IgG 15 days after the first immunization. On the other hand, 60 days after first immunization both adjuvants act benefiting total IgG production similarly. The antibodies of the IgG isotype produced by animals immunized after one or two doses after first immunization, showed intermediate and high avidity, independent on the adjuvant used. In both experimental groups the swelling of the footpads was significantly higher than those of the controls, suggesting that only one dose was enough to stimulate the generation of cellular immunity. Conclusion: The use of this cationic adjuvant for N. meningitidis OMVs preparation revealed good potential for future new antigen preparation for N. meningitidis vaccine. [ABSTRACT FROM AUTHOR]
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- 2021
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4. Study of avidity-ELISA: Comparison of chaotropic agents, incubation temperature and affinity maturation after meningococcal immunization.
- Author
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Portilho, Amanda Izeli, Santos, Jessica Silva, Trzewikoswki de Lima, Gabriela, Lima, Gabrielle Gimenes, and De Gaspari, Elizabeth
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IMMUNIZATION , *ALUMINUM hydroxide , *HUMORAL immunity , *UREA , *ANTIGENS - Abstract
The avidity index (AI) measures the binding strength between the antibody and the antigen, reflecting the affinity maturation. It can be measured by a modified ELISA, adding a chaotropic agent to disrupt the antigen x antibody interaction. However, details of the protocols used affect the final results. We compared the AI of mice sera after a three-dose immunization with meningococcal antigens using different adjuvants. The AI was assessed using potassium thiocyanate (KSCN) and urea as chaotropic agents, incubated at 4 °C, room temperature (RT) and 37 °C. KSCN presented statistically different results when the incubation was set at 4 °C vs RT and 4 °C vs 37 °C, thus, the mean AI obtained were lower. For Urea, 4 °C vs 37 °C presented relevant differences. Using whole-cells suspensions or OMVs as coating antigen provided similar results in some protocols. Thus, the affinity maturation was assessed after each immunization dose and adjuvant use (aluminium hydroxide and dimethyldioctadecylammonium bromide) supported affinity maturation. It is important to study the AI as a functional parameter of humoral response, and both KSCN and Urea are suitable chaotropic agents, however, the protocols should be standardized considering the nature of the antigen, the chaotropic activity and overall laboratory conditions. Adjuvants are important tools to improve antibody avidity following immunization. • Incubation of KSCN or Urea at 4 °C or 37 °C • Correlation of OMVs and WCs were better using KSCN 4 °C/RT or Urea 37 °C. • Affinity maturation curves were similar with different protocols. • Adjuvants improved avidity after immunization in middle-aged mice. [ABSTRACT FROM AUTHOR]
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- 2023
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5. Homologous prime-boost strategy in neonate mice using Neisseria lactamica
- Author
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Ito, André Y., Néri, Simone, Machado, Marta S.S., Tunes, Claudia F., and De Gaspari, Elizabeth N.
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LABORATORY mice , *IMMUNE response , *GRAM-negative bacteria , *BORDETELLA pertussis , *IMMUNOLOGICAL adjuvants , *IMMUNIZATION , *ANTIGENS - Abstract
Abstract: The aim of the present study was to investigate the immune response to native outer membrane vesicles (NOMVs) of Neisseria lactamica with and without Bordetella pertussis (BP) as adjuvant in intranasal (i.n./i.m) immunization. N. lactamica NOMVs delivered intranasally (i.n) to BALB/c mice in a final volume of 5μl that was gradually introduced with a micropipette, Animals received 1, 2, 3, or 4 doses of antigens at 3, 7, 9 and 12 days after birth. On the 35th day, the animals were immunized intramuscularly (i.m.) with (NOMV) of N. lactamica. The prime-booster strategy using NOMV of N. lactamica with BP as adjuvant in the primer (i.n.) and booster (i.m.) is an effective immunization protocol for inducing humoral immune responses producing IgG antibodies of intermediate to high avidity. [Copyright &y& Elsevier]
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- 2009
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6. 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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Ferraz, Aline S., Belo, Elza F. T., Coutinho, Ligia M. C. C., Oliveira, Ana P., Carmo, Andréia M. S., Franco, Daniele L., Ferreira, Tatiane, Yto, André Y., Machado, Marta S. F., Scola, Monica C. G., and De Gaspari, Elizabeth
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IMMUNOGLOBULIN M , *IMMUNOGLOBULIN G , *MONOCLONAL antibodies , *NEISSERIA meningitidis , *POLYSACCHARIDES , *OLIGOSACCHARIDES , *ANTIGENS - Abstract
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]
- Published
- 2008
- Full Text
- View/download PDF
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