title: Immunoaffinity extraction of testosterone by antibody immobilized monolithic capillary with on-line laser-induced fluorescence detection.
abstract: An immunoaffinity capillary column has been made with poly (2-vinyl-4, 4-dimethylazlactone-co-2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) (VDA-co-HEMA-co-EDMA) monolith as a support for the immobilized antibody. The monolith is prepared by UV-initiated in situ polymerization, followed by immobilizing anti-testosterone polyclonal antibody through the rapid reaction with VDA. Fluorescence labeled testosterone at C(3) is designed as a tracer to estimate the extraction ability of this immunoaffinity column, and to optimize the immunoextraction process, such as washing, eluting, incubation and injection. The performance in a more realistic application is then demonstrated successfully for the rapid extraction of testosterone (T) by competitive immunoassay and on-line laser-induced fluorescence (LIF) detection. This immunoaffinity monolith is easily prepared, with high mechanical strength and low flow resistance. It is a satisfactory material as an immunoextractor to detect small molecular compounds specifically and rapidly with high sensitivity (sub ng/mL level).
authors.affiliation: Beijing National Laboratory for Molecular Sciences, BNLMS, Institute of Analytical Chemistry, College of Chemistry, Peking University, Beijing 100871, China.
authors.forename: Hong-Xu
authors.initials: HX
authors.lastname: Chen
authors.forename: Ting
authors.initials: T
authors.lastname: Huang
authors.forename: Xin-Xiang
authors.initials: XX
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D007118:Immunoassay; D007164:Immunosorbents; D011108:Polymers / Q000737:chemistry; D013739:Testosterone / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000906:Antibodies; D007164:Immunosorbents; D011108:Polymers; D013739:Testosterone