Desenvolvimento e caracterização de imunosorventes e avaliação de desempenho em ensaios imunoenzimaticos

AUTOR(ES)

Leila Peres

DATA DE PUBLICAÇÃO

1986

RESUMO

In this work the characterization of a new type of support for the immobilization of antigens and antibodies was initiated. This support that was firstly described by Bittencourt et al in 1983, consists of synthetic fabrics coated with crosslinked resins containing N-methylol groups able to form covalent bonds, with functional groups such as hydroxil and amino, which are characteristic of proteins. Immunosorbents prepared with characteristic antigens of different pathologies, such as Chagas Disease and Toxoplasmosis (parasitic diseases) and Rubella (viruse), presented excellent performance in sorologic diagnosis, when utilized in ELISA tests (Enzyme-Linked lmmunosorbent Assays). The studies conducted suggest that the immobilization reaction, is controlled by the surface area available, associated to the area occupied by the molecule. The results indicate, covalent bonding of chemical adsorption, of the immunoproteins with N-methylol groups, which are present in excess at the surface of the crosslinked resin. This first layer seems to be followed by multilayers sorbed by weaker forces. The Adsorption Isotherm obtained for the immobilization of human antibody, IgG, presented a sigmoidal form corresponding to models, such as B.E.T. and others, which assume the formation of multilayers. With respect to polystirene surfaces, which are presently the most utilized type of support in ELISA tests, the isotherms obtained were fitted by the Langmuir Model. This same model was consistent, with the isotherms obtained from the capture of the human antibody IgG in solution, by Anti-IgG, previously immobilized on the supports, demonstrating monolayer adsorption. Relevant variables for the immobilization reaction, such as: pH, temperature, time of reaction, and concentrations of immunoproteins and electrolytes were studied, with the objective of determining the most adequated reaction conditions. A systematic study of the performance of the immunosorbent developed in this work, was conducted with ELISA tests for Chagas` Disease, showing the same levels of sensitivity, specificity and reproductibility of polystirene plates, with the advantage of superior stability.

ASSUNTO(S)

imunoquimica materiais

Documentos Relacionados

• Vírus da artrite encefalite caprina:: desenvolvimento e padronização de ensaios imunoenzimáticos (ELISA e Dot-Blot) e estudo epidemiológico no Estado do Ceará
• Avaliação de frações hidrofóbicas e hidrofílicas de brucella abortus em ensaios imunoenzimáticos para caracterizar o perfil de anticorpos produzidos por bovinos vacinados e não-vacinados
• AvaliaÃÃo de fraÃÃes hidrofÃbicas e hidrofÃlicas de brucella abortus em ensaios imunoenzimÃticos para caracterizar o perfil de anticorpos produzidos por bovinos vacinados e nÃo-vacinados
• Comparação entre ensaios imunoenzimáticos realizados em amostras de sangue seco e soro para triagem prénatal da toxoplasmose: Estudo populacional
• Desenvolvimento e avaliação de um sistema automatizado para ensaios de molinetes