Please use this identifier to cite or link to this item:
https://www.arca.fiocruz.br/handle/icict/18187
FLOW CYTOMETRY AS A TOOL FOR QUALITY CONTROL OF FLUORESCENT CONJUGATES USED IN IMMUNOASSAYS
Controle de qualidade
Imunoensaios
Conjugados Fluorescentes
Author
Affilliation
Fundação Oswaldo Cruz. Instituto de Tecnologia Imunobiológica - Farmanguinhos. Laboratório de Tecnologia de Diagnóstico. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto de Tecnologia Imunobiológica - Farmanguinhos. Laboratório de Tecnologia de Diagnóstico. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto de Tecnologia Imunobiológica - Farmanguinhos. Laboratório de Tecnologia de Diagnóstico. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto de Tecnologia Imunobiológica - Farmanguinhos. Laboratório de Tecnologia de Diagnóstico. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunoparasitologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunologia Clínica. Rio de Janeiro, RJ. Brasil.
Fundação Oswaldo Cruz. Instituto de Tecnologia Imunobiológica - Farmanguinhos. Laboratório de Tecnologia de Diagnóstico. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto de Tecnologia Imunobiológica - Farmanguinhos. Laboratório de Tecnologia de Diagnóstico. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto de Tecnologia Imunobiológica - Farmanguinhos. Laboratório de Tecnologia de Diagnóstico. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunoparasitologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunologia Clínica. Rio de Janeiro, RJ. Brasil.
Abstract
The use of antibodies in immunodiagnostic kits generally implies the conjugation of these proteins with other molecules such as chromophores or fluorochromes. The development of more sensitive quality control procedures than spectrophotometry is essential to assure the use of better fluorescent conjugates since the fluorescent conjugates are critical reagents for a variety of immunodiagnostic kits. In this article, we demonstrate a new flow cytometric protocol to evaluate conjugates by molecules of equivalent soluble fluorochromes (MESF) and by traditional flow cytometric analysis. We have coupled microspheres with anti-IgG-PE and anti-HBSAg-PE conjugates from distinct manufactures and/or different lots and evaluated by flow cytometry. Their fluorescence intensities were followed for a period of 18 months. Our results showed that there was a great difference in the fluorescence intensities between the conjugates studied. The differences were observed between manufactures and lots from both anti-IgG-PE and anti-HBSAg-PE conjugates. Coefficients of variation (CVs) showed that this parameter can be used to determine better coupling conditions, such as homogenous coupling. The MESF analysis, as well as geometric mean evaluation by traditional flow cytometry, showed a decrease in the values for all conjugates during the study and were indispensable tools to validate the results of stability tests. Our data demonstrated the feasibility of the flow cytometric method as a standard quality control of immunoassay kits.
Keywords in Portuguese
Citometria de flucoControle de qualidade
Imunoensaios
Conjugados Fluorescentes
Share