title: Development of a Fluorescence-Linked Immunosorbent Assay for Baicalin.
abstract: Previously, we developed an indirect competitive enzyme-linked immunosorbent assay (icELISA) for baicalin (BAL) and used this assay to investigate the pharmacokinetics of BAL in mice. In this study, an anti-BAL monoclonal antibody (MAb) was purified by the caprylic acid method and then labelled with fluorescein isothiocyanate (FITC). Subsequently, an indirect competitive fluorescence-linked immunosorbent assay (icFLISA) was developed to detect baicalin (BAL) using FITC-labelled anti-BAL MAbs. Characterization of the assay demonstrated an effective BAL measurement range of 6.4 ng/mL to 500 μg/mL (R(2) = 0.997). The relative standard deviations (RSDs) for both intra-assay and inter-assay repeatability and precision were below 10 %. This icFLISA for BAL is simple, rapid and sensitive, with a 390-fold larger linear range and a 2-fold lower limit of detection (LOD) compared with the previously developed icELISA. We observed a strong correlation between the results determined by the icFLISA and icELISA methods. Overall, this study provides a useful method for detecting BAL in medicines, enabling in vivo visualization research.
authors.affiliation: School of Chinese Materia Medica, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China.
authors.forename: Wenchao
authors.initials: W
authors.lastname: Shan
authors.affiliation: School of Chinese Materia Medica, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China.
authors.forename: Jinjun
authors.initials: J
authors.lastname: Cheng
authors.affiliation: School of Basic Medical Sciences, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China.
authors.forename: Baoping
authors.initials: B
authors.lastname: Qu
authors.affiliation: The third affiliated hospital, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China.
authors.forename: Jiayang
authors.initials: J
authors.lastname: Sai
authors.affiliation: School of Basic Medical Sciences, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Kong
authors.affiliation: Center of Scientific Experiment, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China. quhuihua@gmail.com.
authors.forename: Huihua
authors.initials: H
authors.lastname: Qu
authors.affiliation: School of Basic Medical Sciences, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: School of Basic Medical Sciences, Beijing University of Chinese Medicine, 11 Beisanhuandong Road, Chaoyang District, Beijing, 100029, China.
authors.forename: Qingguo
authors.initials: Q
authors.lastname: Wang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D005419:Flavonoids / Q000032:analysis* / Q000276:immunology; D007163:Immunosorbent Techniques; D051379:Mice; D013050:Spectrometry, Fluorescence
chemical_list: D000911:Antibodies, Monoclonal; D005419:Flavonoids; C038044:baicalin
keywords: Baicalin; Enzyme-linked immunosorbent assay; Fluorescence-linked immunosorbent assay; Fluorescently labelled monoclonal antibody; Traditional Chinese medicine