Development and application of a multi-target immunoaffinity column for the selective extraction of natural estrogens from pregnant women's urine samples by capillary electrophoresis.
In this paper, a methodology for the determination of three naturally occurring estrogens (estradiol, estrone and estriol) in pregnant women's urine has been described. The procedure included immunoaffinity column (IAC) extraction of 4 mL of urine sample and subsequent analysis of the extraction by micellar electrokinetic chromatography (MEKC). A multi-target polyclonal antibody that has high affinity to three estrogens was produced. Then the IAC was developed by coupling polyclonal antibody to CNBr-activated Sepharose 4B. The IAC showed high affinity for these estrogens. Recoveries of three estrogens from human serum matrix were greater than 92% with R.S.D. less than 4.5%. The final elute of urine sample was diluted with running buffer and then quantitated with MEKC. The experimental results demonstrated that IAC was a useful technique for extraction and concentration of estrogens from biological samples. Three estrogens levels in six pregnant women's urine were measured by both the present method and enzyme-linked immunoadsorbent assay (ELISA). The results of this method have been found to correlate well with those of ELISA.
Su
Ping
P
College of Chemistry, Peking University, Beijing 100871, China.
Zhang
Xin-Xiang
XX
Chang
Wen-Bao
WB
D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D020374:Chromatography, Micellar Electrokinetic Capillary; D004958:Estradiol / Q000302:isolation & purification / Q000652:urine; D004964:Estriol / Q000302:isolation & purification / Q000652:urine; D004967:Estrogens / Q000302:isolation & purification* / Q000652:urine*; D004970:Estrone / Q000302:isolation & purification / Q000652:urine; D005260:Female; D006801:Humans; D011247:Pregnancy
D004967:Estrogens; D004970:Estrone; D004958:Estradiol; D004964:Estriol