2023 Volume 72 Issue 4.5 Pages 167-173
We have developed a new quantitative method for polysaccharide by sandwich enzyme-linked lectin assay (ELLA). In this high specificity method, polysaccharide is reacted with lectin immobilized in a 96-well microtiter plate, and then is sandwiched by enzyme-conjugated lectin, followed by color development by the enzyme activities. Guar gam was detected at a concentration from 0.1 μg mL−1 to 3 μg mL−1, and exopolysaccharide (EPS) from Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1 (R-1) was detected at a concentration from 0.2 μg mL−1 to 1 μg mL−1 by this method. We quantified EPS in the culture fluid of R-1 strain after removing proteins by this method. The values obtained in spike and recovery test were from 97 % to 99 %. The precision of analysis was that repeatability was 6.2 % and intermediate precision was 6.8 %. These results showed this method had enough analytical ability. Furthermore, due to good operability, this method is expected to be applied to the quantification of various polysaccharides.
