title: Development of immunoaffinity chromatography to specifically knockout baicalin from Gegenqinlian Decoction.
abstract: Specific knockout technology provides a powerful tool to confirm the role of target compounds in a plant or its derived prescriptions, and this principle is the same as that with knockout genes. In this study, we generated an immunoaffinity column conjugated with an anti-baicalin monoclonal antibody and then loaded Gegenqinlian Decoction extracts, followed by washing with deionized water and an elution solvent. The results of the high-performance liquid chromatography fingerprints and high-performance liquid chromatography with mass spectrometry showed that the immunoaffinity column was able to specifically knockout baicalin, oroxylin A-7-O-glucuronide, wogonoside, wogonin, and baicalein from Gegenqinlian Decoction. A reliable one-step method to specifically knockout baicalin was established with an immunoaffinity column. Gegenqinlian Decoction and its knocked-out fraction induced the expression of superoxide dismutase and were compared in human umbilical vein endothelial cells cultured with a high glucose concentration; the results showed that the Gegenqinlian Decoction and its knocked-out fraction showed no significant difference, which indicated that the baicalin, oroxylin A-7-O-glucuronide, wogonoside, wogonin, and baicalein that were knocked out by the immunoaffinity column might not be key compounds for the induction of Gegenqinlian Decoction superoxide dismutase secretion.
authors.affiliation: School of Basic Medical Sciences, Beijing University of Chinese Medicine, Chaoyang District, Beijing, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: School of Chinese Materia Medica, Beijing University of Chinese Medicine, Chaoyang District, Beijing, China.
authors.forename: Huibin
authors.initials: H
authors.lastname: Feng
authors.affiliation: School of Chinese Materia Medica, Beijing University of Chinese Medicine, Chaoyang District, Beijing, China.
authors.forename: Wenchao
authors.initials: W
authors.lastname: Shan
authors.affiliation: School of Chinese Materia Medica, Beijing University of Chinese Medicine, Chaoyang District, Beijing, China.
authors.forename: Jinjun
authors.initials: J
authors.lastname: Cheng
authors.affiliation: School of Basic Medical Sciences, Beijing University of Chinese Medicine, Chaoyang District, Beijing, China.
authors.forename: Xueqian
authors.initials: X
authors.lastname: Wang
authors.affiliation: School of Basic Medical Sciences, Beijing University of Chinese Medicine, Chaoyang District, Beijing, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Scientific Research Experiment Center, Beijing University of Chinese Medicine, Chaoyang District, Beijing, China.
authors.forename: Huihua
authors.initials: H
authors.lastname: Qu
authors.affiliation: School of Basic Medical Sciences, Beijing University of Chinese Medicine, Chaoyang District, Beijing, China.
authors.forename: Qingguo
authors.initials: Q
authors.lastname: Wang
keywords: Baicalin; Gegenqinlian Decoction; Immunoaffinity columns; Monoclonal antibodies; Superoxide dismutase