title: Development of a monoclonal antibody-based enzyme-linked immunosorbent assay for luteoloside detection in Flos Lonicerae Japonicae.
abstract: Flos Lonicerae Japonicae (FLJ), the flower bud of Lonicera japonica Thunb. (Caprifoliaceae), is a widely used traditional Chinese medicine with various pharmacological activities. Luteoloside is a major active compound and a quality control marker of FLJ. Luteolin-7-O-glucuronide (LG), an analog of luteoloside, was conjugated with bovine serum albumin (BSA) and ovalbumin (OVA) to create the immunogen and coating antigen, respectively. A sensitive and specific monoclonal antibody (mAb), designated as mAb3A4, was generated with LG-BSA. To screen the authenticity and quality of FLJ, an indirect competitive enzyme-linked immunosorbent assay (icELISA) was established. The concentration of luteoloside producing 50 % inhibition and the working range of the icELISA were 42.3 and 9.1-258.1 μg L(-1), respectively. The icELISA showed cross-reactivity values of 2414, 402, 230, and <1 % for LG, baicalin, scutellarin, and other analogs of luteoloside, respectively. The average recovery of luteoloside in the FLJ samples as determined by icELISA ranged from 83.0 to 112.5 %. The luteoloside content was determined for different Lonicera herbal samples with icELISA, and the results were confirmed by high-performance liquid chromatography analysis. Thus, this icELISA is suitable for the quality assurance of FLJ samples. Graphical abstract Specific monoclonal antibody-based enzyme-linked immunosorbent assay for luteoloside.
authors.affiliation: State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.|College of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan, Shandong, 250355, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Zhang
authors.affiliation: State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors.forename: Tiegui
authors.initials: T
authors.lastname: Nan
authors.affiliation: State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors.forename: Zhilai
authors.initials: Z
authors.lastname: Zhan
authors.affiliation: State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors.forename: Liping
authors.initials: L
authors.lastname: Kang
authors.affiliation: State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Yang
authors.affiliation: State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: College of Agriculture and Biotechnology, China Agricultural University, Beijing, 100193, China. wbaomin@263.net.
authors.forename: Baomin
authors.initials: B
authors.lastname: Wang
authors.affiliation: State Key Laboratory Breeding Base of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, 100700, China. huangluqi01@126.com.
authors.forename: Luqi
authors.initials: L
authors.lastname: Huang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D000917:Antibody Formation; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D005960:Glucosides / Q000032:analysis* / Q000276:immunology*; D006241:Haptens / Q000737:chemistry / Q000276:immunology; D031239:Lonicera / Q000737:chemistry* / Q000276:immunology; D047311:Luteolin / Q000032:analysis* / Q000737:chemistry / Q000276:immunology*; D008807:Mice, Inbred BALB C; D010936:Plant Extracts / Q000737:chemistry* / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D005960:Glucosides; D006241:Haptens; D010936:Plant Extracts; C456096:luteolin-7-O-glucuronide; C588038:lonicerae flos; C066408:luteolin-7-glucoside; D047311:Luteolin
keywords: ELISA; Flos Lonicerae Japonicae; Luteoloside; Monoclonal antibody; Quality control