An ELISA-based Test System for the KombiVIChvak Anti-HIV Vaccine Control during its Production

Автор: O.N. Kaplina, L.I. Karpenko, V.S. Kaplin, L.R. Lebedev, A.A. Ilichev, and M.P. Bogryantseva

Страница: 85-93

An ELISA-based Test System for the KombiVIChvak Anti-HIV Vaccine Control during its Production

Biotekhnologiya, 2015, N 4, P. 85-93

UDC 617.017: 615.371: 577.1

Section:  “Metrology, Standardization, and Control”

O.N. Kaplina ^*,  L.I. Karpenko,  V.S. Kaplin,  L.R. Lebedev,  A.A. Ilichev,  and  M.P. Bogryantseva

State Research Center for Virology and Biotechnology «Vector»,  630559, Kol'tsovo, Novosibirskaya oblast Russia

e-mail:  okaplina@vector.nsc.ru,  forelat@ngs.ru

A test system to assess the activity of the KombiVIChvak anti-HIV-1 vaccine has been designed. Standard panels of negative and positive samples of murine sera were created. The conditions for the analysis using a polyepitope recombinant protein TBI were optimized. The virus-neutralizing activity of the vaccine was assessed in sera of immunized human and mice, and a high specificity and sensitivity of the designed test-system was demonstrated. The results obtained using this system were confirmed in the assay with the NewLavBlot analytical set; it was shown that all the samples that were identified as positive by the suggested test system contained the HIV-1characteristic proteins. The validation was performed; the intra- and inter-laboratory accuracies of the designed system were determined. The reproducibility of the system was equal to 2—10% with the satisfactory value equal to 19%.

Key words:  antibodies,  anti-HIV-1 vaccine,  ELISA-based test system,  polyepitope TBI protein,  sensitivity,  specificity,  validation.

06.11.2015, 1662 просмотра.