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. 2017 Aug 4;7(1):7288.
doi: 10.1038/s41598-017-07687-7.

A time-resolved fluoroimmunoassay to assay the rabies virus glycoprotein: application for estimation of human rabies vaccine potency

Guanfeng Lin  1 , Shaolang Chen  2 , Hui Zhao  3 , Junyu Liang  2 , Qiaoting Deng  2 , Rongliang Liang  2 , Xinxin Guo  2 , Zhenhua Chen  2 , Baihong Chen  1 , Tiancai Liu  2   4 , Yingsong Wu  5   6
Affiliations

A time-resolved fluoroimmunoassay to assay the rabies virus glycoprotein: application for estimation of human rabies vaccine potency

Guanfeng Lin et al. Sci Rep. .

Abstract

Replacement of the in vivo rabies vaccine potency test (NIH test) by in vitro methods had been discussed by several researcher including WHO expert working groups. In this paper, a time-resolved fluoroimmunoassay (TRFIA) for the assay of rabies virus glycoprotein in rabies vaccine was first established to estimate the rabies vaccine potency by using specific monoclonal antibody that only recognized the native, trimeric and immunogenic form of rabies virus glycoprotein. Potency of the rabies virus glycoprotein was assayed with satisfactory performance under optimal conditions, and the method demonstrated satisfactory results when applied in practical samples. The correlation coefficient of potency values obtained from the present TRFIA and ELISA was 0.912, and 0.903 for those from the present TRFIA and NIH test. These preliminary results confirmed that this TRFIA can replace ELISA with higher performance, and could be a promising replacement of the NIH test. Based upon these results, the present TRFIA seemed to be a convenient tool for evaluating rabies vaccine potency and its products at different stages accordingly.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Example of the present TRFIA employing a europium chelate label.
Figure 2
Figure 2
Standard curves and intra-assay precision profile for the present TRFIA (each point was based on 10 replicates).
Figure 3
Figure 3
High-dose signal saturation (hook-effect) for the present TRFIA (each point was based on 10 replicates).
Figure 4
Figure 4
Graphical comparisons of the present TRFIA and ELISA results for assay of rabies virus glycoprotein.
Figure 5
Figure 5
Result of TRFIA and NIH method for thirty nine rabies vaccine samples.
Figure 6
Figure 6
Graphical comparisons of the present TRFIA and NIH test results for evaluation of the potency of rabies vaccine samples.
See this image and copyright information in PMC

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