Key result
A novel GloSensor-based reporter assay for rhBNP demonstrated high precision, accuracy, and linearity, with no significant difference in test results compared to a traditional ELISA assay (p=0.630).
A novel GloSensor-based reporter assay provides a rapid, simple, and accurate method for determining the biological activity of recombinant human brain natriuretic peptide (rhBNP).
May enable alternative rhBNP quantification in labs; leaves open clinical adoption pending further validation.
Accurate determination of biological activity is essential in quality control of recombinant human brain natriuretic peptide (rhBNP). In previous study, we successfully developed a genetically modified cell line 293GCAC3-based ELISA assay for rhBNP. But ELISA procedure is still tedious, so this study was aimed to develop a rapid and simple bioassay for rhBNP using GloSensor technology, which provides a platform of flexible luciferase-based biosensors for real-time detection of signaling events in live cells, including cGMP production. A reporter cell line 293GCAGlo-G1 was constructed by transfecting pGloSensor™ 40 F plasmid into 293GCAC3. The reporter assay based on 293GCAGlo-G1 showed high precision with intra-assay CV being 8.3% and inter-assay CV being 14.1%; high accuracy with 80%, 100% and 120% recovery rate being 99.2%, 102.4% and 99.0% respectively; and great linearity with R 2 of linear fitting equation being 0.99. Besides, no significant difference was found in test results of reporter assay and 293GCAC3-based ELISA assay (paired t test, p = 0.630). All these results suggested that the reporter assay was a viable assay for biological determination of rhBNP.
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Yu et al. (2018) studied Quality control of recombinant human brain natriuretic peptide (rhBNP). GloSensor-based reporter assay (293GCAGlo-G1) vs. 293GCAC3-based ELISA assay was evaluated on Test results comparison between reporter assay and ELISA assay (p=0.630). A novel GloSensor-based reporter assay for rhBNP demonstrated high precision, accuracy, and linearity, with no significant difference in test results compared to a traditional ELISA assay (p=0.630).
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