Immunoaffinity extraction of testosterone by antibody immobilized monolithic capillary with on-line laser-induced fluorescence detection.
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).
Chen
Hong-Xu
HX
Beijing National Laboratory for Molecular Sciences, BNLMS, Institute of Analytical Chemistry, College of Chemistry, Peking University, Beijing 100871, China.
Huang
Ting
T
Zhang
Xin-Xiang
XX
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*
D000906:Antibodies; D007164:Immunosorbents; D011108:Polymers; D013739:Testosterone