title: Development of an ELISA and immunochromatographic strip for highly sensitive detection of microcystin-LR.
abstract: A monoclonal antibody for microcystin-leucine-arginine (MC-LR) was produced by cell fusion. The immunogen was synthesized in two steps. First, ovalbumin/ bovine serum albumin was conjugated with 6-acetylthiohexanoic acid using a carbodiimide EDC (1-ethyl-3-[3-dimethylaminopropyl]carbodiimide hydrochloride)/ NHS (N-hydroxysulfosuccinimide) reaction. After dialysis, the protein was reacted with MC-LR based on a free radical reaction under basic solution conditions. The protein conjugate was used for immunization based on low volume. The antibodies were identified by indirect competitive (ic)ELISA and were subjected to tap water and lake water analysis. The concentration causing 50% inhibition of binding of MC-LR (IC50) by the competitive indirect ELISA was 0.27 ng/mL. Cross-reactivity to the MC-RR, MC-YR and MC-WR was good. The tap water and lake water matrices had no effect on the detection limit. The analytical recovery of MC-LR in the water samples in the icELISA was 94%-110%. Based on this antibody, an immunochromatographic biosensor was developed with a cut-off value of 1 ng/mL, which could satisfy the requirement of the World Health Organization for MC-LR detection in drinking water. This biosensor could be therefore be used as a fast screening tool in the field detection of MC-LR.
authors.affiliation: State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, JiangSu 214122, China. raxray@gmail.com.
authors.forename: Liqiang
authors.initials: L
authors.lastname: Liu
authors.affiliation: State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, JiangSu 214122, China. da_rui12345@163.com.
authors.forename: Changrui
authors.initials: C
authors.lastname: Xing
authors.affiliation: State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, JiangSu 214122, China. yanhuijuan7@163.com.
authors.forename: Huijuan
authors.initials: H
authors.lastname: Yan
authors.affiliation: State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, JiangSu 214122, China. kuangh@jiangnnan.edu.cn.
authors.forename: Hua
authors.initials: H
authors.lastname: Kuang
authors.affiliation: State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, JiangSu 214122, China. da_rui12345@163.com.
authors.forename: Chuanlai
authors.initials: C
authors.lastname: Xu
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry; D001120:Arginine / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007930:Leucine / Q000737:chemistry*; D057230:Limit of Detection; D052998:Microcystins / Q000737:chemistry*; D011934:Reagent Strips / Q000737:chemistry*
chemical_list: D000911:Antibodies, Monoclonal; D052998:Microcystins; D011934:Reagent Strips; C078588:microcystin; D001120:Arginine; D007930:Leucine