ABSTRACT
@#To synthesize the folic acid-alliinase conjugate(FA-Alliinase), and to study its targeting and antitumor activity on cervical cancer HeLa cells. FA-Alliinase I and FA-Alliinase II were synthesized by two methods. The couping ratios of two conjugates measured were 12 and 31, respectively. The FA-Alliinase II with high coupling ratio was selected and its structure was characterized preliminarily. The activity of alliinase retained about 50% in FA-Alliinase II determined by HPLC. The specific effect of FA-Alliinase II on HeLa cells was observed by confocal laser and flow cytometry. The antitumor activity of conjugate combined with alliin was determined by MTT, and IC50 of alliin was(127. 6±2. 3)μmol/L. This study provides a direct evaluation method for the synthesis and optimization of FA-Alliinase.
ABSTRACT
Hapten-carrier protein conjugates were made using ciprofloxacin (CPFX) and two carrier proteins by 1-ethyl-3-(3- dimethylaminopropyl)-carbodiimide hydrochloride (EDC) method. Ultraviolet spectrophotometry were used to demonstrated that the molecule conjugate ratio of CPFX to ovalbumin (OVA) and bovine serum albumin (BSA) are 6:1 and 13:1 respectively. Nondenaturing gel electrophoresis results revealed that the conjugate band migrates differently from that of the carrier protein alone and of the EDC-treated protein when as few as 6 molecules of CPFX are attached to the carrier protein. The results indicate that nondenaturing gel electrophoresis and ultraviolet spectrophotometry can be employed to analyze the molecule coupling ratio of CPFX to carrier proteins qualitatively and quantitatively.