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Received: 2004-03-02

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Journal of Zhejiang University SCIENCE A 2004 Vol.5 No.8 P.900~905


Affinity ultrafiltration of DNA topoisomerases-targeted compounds determined with HPLC/ESI-MS for drug candidate screening

Author(s):  ZHANG Hong, PAN Yuan-jiang

Affiliation(s):  Department of Chemistry, Zhejiang University, Hangzhou 310027, China; more

Corresponding email(s):   panyuanjiang@css.zju.edu.cn

Key Words:  Affinity, High-throughput screening, LC-MS, Topoisomerase, Ultrafiltration

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ZHANG Hong, PAN Yuan-jiang. Affinity ultrafiltration of DNA topoisomerases-targeted compounds determined with HPLC/ESI-MS for drug candidate screening[J]. Journal of Zhejiang University Science A, 2004, 5(8): 900~905.

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%T Affinity ultrafiltration of DNA topoisomerases-targeted compounds determined with HPLC/ESI-MS for drug candidate screening
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T1 - Affinity ultrafiltration of DNA topoisomerases-targeted compounds determined with HPLC/ESI-MS for drug candidate screening
A1 - ZHANG Hong
A1 - PAN Yuan-jiang
J0 - Journal of Zhejiang University Science A
VL - 5
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SP - 900
EP - 905
%@ 1869-1951
Y1 - 2004
PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.2004.0900

A method of screening assay is demonstrated. The approach is based on the affinity of antitumor candidates for topoi-somerases. In this method, antitumor candidates are fished out using topoisomerases as targets. Traditional analysis of complex compounds typically encounters signal suppression due to the relatively low concentrations, but enzyme-affinity screening for the active compounds can effectively concentrate the desired analysts into a small volume of high concentration. Active compounds are separated from non-affinity compounds by ultrafiltration. The molecules-enzymes complexes that are retained on the filter are subsequently separated by acidification to obtain the topoisomerases-affinity compounds for analysis on High Performance Liquid Chromatography coupled with electrospray ionization mass spectrometric detection (ESI-MS). This enzyme-affinity based screening assay provides a highly specific and efficient method that can directly screen, identify, and acquire drug candidates thus improving the accuracy and speed of high-throughput screening activities.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article


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