A rapid reporter assay for recombinant human brain natriuretic peptide (rhBNP) by GloSensor technology
- Author:
Lei YU
1
;
Xinchang SHI
;
Chunmei HAN
;
Chunming RAO
;
Junzhi WANG
Author Information
1. National Institutes for Food and Drug Control
- Keywords:
RhBNP;
cGMP;
GloSensor technology;
Reporter assay
- From:
Journal of Pharmaceutical Analysis
2018;8(5):297-301
- CountryChina
- Language:Chinese
-
Abstract:
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 R2of 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.
