Optimization research on in vitro screening model for human leukocyte elastase inhibitor
10.3724/SP.J.1008.2011.00640
- Author:
Ge SONG
1
Author Information
1. Department of Biochemistry and Molecular Biology
- Publication Type:Journal Article
- Keywords:
Inhibitor;
Leuksocyte elastase;
Reaction condition optimization;
Screening model
- From:
Academic Journal of Second Military Medical University
2011;32(6):640-644
- CountryChina
- Language:Chinese
-
Abstract:
Objeciive To explore the most suitable reaction condition for human leukocyte elastase (HLE) and its substrates, so as to establish a high-throuthput in vitro screercng model for HLE mhibiton Methods By measuring the UV absorbance of the product p-NA, we studied the dectrolyte concentration, temperature, pH, enzyme/substrate concentration, the influence of reaction time, and deteclion wavelength on the reaclion, so as to determine the optimal condilion for the reaclion between HLE and its substrates. Results The suitable reaction condition was determined as followes: HLE (0. 2 U/ml) 30 μl, substrates (50 mmol/L) 100 μl, Ttiis buffer (1.0 mol/L NaCl, pH 7. 5), temperatiire 37°C, reaction lime 24 h, and detection wavelength 545 nm. Conclution We have successfully established a stable in vitfo HLE inhibitor screening model with high sensitivity, accuracy, and repeatability, which provides a better platform for HLE inhibitor screening.
