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Flow Cytometry as a Tool for Quality Control of Fluorescent Conjugates Used in Immunoassays.
de Almeida Santiago, Marta; de Paula Fonseca E Fonseca, Bruna; da Silva Marques, Christiane de Fátima; Domingos da Silva, Edimilson; Bertho, Alvaro Luiz; Nogueira, Ana Cristina Martins de Almeida.
Afiliação
  • de Almeida Santiago M; Laboratory of Diagnostic Technology, Immunobiological Technology Institute, FIOCRUZ, Rio de Janeiro, Rio de Janeiro, Brazil.
  • de Paula Fonseca E Fonseca B; Laboratory of Diagnostic Technology, Immunobiological Technology Institute, FIOCRUZ, Rio de Janeiro, Rio de Janeiro, Brazil.
  • da Silva Marques CF; Laboratory of Diagnostic Technology, Immunobiological Technology Institute, FIOCRUZ, Rio de Janeiro, Rio de Janeiro, Brazil.
  • Domingos da Silva E; Laboratory of Diagnostic Technology, Immunobiological Technology Institute, FIOCRUZ, Rio de Janeiro, Rio de Janeiro, Brazil.
  • Bertho AL; Laboratory of Immunoparasitology, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Rio de Janeiro, Brazil.
  • Nogueira AC; Flow Cytometry Core Facility, Oswaldo Cruz Institute, FIOCRUZ, Rio de Janeiro, Rio de Janeiro, Brazil.
PLoS One ; 11(12): e0167669, 2016.
Article em En | MEDLINE | ID: mdl-27936034
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Imunoensaio / Imunoconjugados / Anticorpos Imobilizados / Citometria de Fluxo / Corantes Fluorescentes Limite: Animals / Humans Idioma: En Revista: PLoS One Assunto da revista: CIENCIA / MEDICINA Ano de publicação: 2016 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Imunoensaio / Imunoconjugados / Anticorpos Imobilizados / Citometria de Fluxo / Corantes Fluorescentes Limite: Animals / Humans Idioma: En Revista: PLoS One Assunto da revista: CIENCIA / MEDICINA Ano de publicação: 2016 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos