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Determination of doping methyltestosterone by capillary electrophoresis immunological analysis with thermally reversible hydrogel and laser-induced fluorescence.
Electrophoresis. 1999 Jul; 20(10):1998-2002.E

Abstract

In this paper thermally reversible hydrogel used as a replaceable packed material for capillary electrophoresis was examined. A simple and rapid method of detecting doping methyltestosterone (MTS) was developed, which demonstrated the potential of strong affinity antibodies (k(d) = 10(9)) as a selector for immunologically based separations in serum, not in urine, by capillary electrophoresis. Polyclonal antibodies (Ab) were polymerized on the hydrogel and applied in the separation of fluorescein isothiocyanate (FITC)-labeled antigen and free antigen with laser-induced fluorescence detector (LIF). The results indicated that N-isopropylacrylamide (PNIPA) hydrogel is a steady, replaceable gel. The specific determination of MTS did not require reaching equilibrium of immunological reaction and the PNIPA hydrogel including antibodies can be stockpiled at 4 degrees C, in preparation for determining MTS at any time. It can be used to determine MTS with good precision with a sensitivity lower than 50 ng/mL. The theoretical plate numbers obtained could reach 168,449 for free-antigen. Details of the preparation of hydrogel cross-linked polyclonal antibody and of typical separations of bound and free antigen are presented.

Authors+Show Affiliations

Department of Chemistry, Peking University, Beijing, China. zxx@chem.pku.edu.cnNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

10451108

Citation

Zhang, X X., et al. "Determination of Doping Methyltestosterone By Capillary Electrophoresis Immunological Analysis With Thermally Reversible Hydrogel and Laser-induced Fluorescence." Electrophoresis, vol. 20, no. 10, 1999, pp. 1998-2002.
Zhang XX, Li J, Gao J, et al. Determination of doping methyltestosterone by capillary electrophoresis immunological analysis with thermally reversible hydrogel and laser-induced fluorescence. Electrophoresis. 1999;20(10):1998-2002.
Zhang, X. X., Li, J., Gao, J., Sun, L., & Chang, W. B. (1999). Determination of doping methyltestosterone by capillary electrophoresis immunological analysis with thermally reversible hydrogel and laser-induced fluorescence. Electrophoresis, 20(10), 1998-2002.
Zhang XX, et al. Determination of Doping Methyltestosterone By Capillary Electrophoresis Immunological Analysis With Thermally Reversible Hydrogel and Laser-induced Fluorescence. Electrophoresis. 1999;20(10):1998-2002. PubMed PMID: 10451108.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Determination of doping methyltestosterone by capillary electrophoresis immunological analysis with thermally reversible hydrogel and laser-induced fluorescence. AU - Zhang,X X, AU - Li,J, AU - Gao,J, AU - Sun,L, AU - Chang,W B, PY - 1999/8/18/pubmed PY - 2000/8/12/medline PY - 1999/8/18/entrez SP - 1998 EP - 2002 JF - Electrophoresis JO - Electrophoresis VL - 20 IS - 10 N2 - In this paper thermally reversible hydrogel used as a replaceable packed material for capillary electrophoresis was examined. A simple and rapid method of detecting doping methyltestosterone (MTS) was developed, which demonstrated the potential of strong affinity antibodies (k(d) = 10(9)) as a selector for immunologically based separations in serum, not in urine, by capillary electrophoresis. Polyclonal antibodies (Ab) were polymerized on the hydrogel and applied in the separation of fluorescein isothiocyanate (FITC)-labeled antigen and free antigen with laser-induced fluorescence detector (LIF). The results indicated that N-isopropylacrylamide (PNIPA) hydrogel is a steady, replaceable gel. The specific determination of MTS did not require reaching equilibrium of immunological reaction and the PNIPA hydrogel including antibodies can be stockpiled at 4 degrees C, in preparation for determining MTS at any time. It can be used to determine MTS with good precision with a sensitivity lower than 50 ng/mL. The theoretical plate numbers obtained could reach 168,449 for free-antigen. Details of the preparation of hydrogel cross-linked polyclonal antibody and of typical separations of bound and free antigen are presented. SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/10451108/Determination_of_doping_methyltestosterone_by_capillary_electrophoresis_immunological_analysis_with_thermally_reversible_hydrogel_and_laser_induced_fluorescence_ L2 - https://doi.org/10.1002/(SICI)1522-2683(19990701)20:10<1998::AID-ELPS1998>3.0.CO;2-C DB - PRIME DP - Unbound Medicine ER -