Development of a New Quantitative Method for Polysaccharide by Sandwich Enzyme-Linked Lectin Assay Using Single Lectin Species

Development of a New Quantitative Method for Polysaccharide by Sandwich Enzyme-Linked Lectin Assay Using Single Lectin Species

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 develo...

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Veröffentlicht in:BUNSEKI KAGAKU 2023/04/05, Vol.72(4.5), pp.167-173
Hauptverfasser: OSHIMA, Toshifumi, NAKAYAMA, Kai, TAKABAYASHI, Kumiko, IWAKI, Misato, UBE, Nana, NAKAJIMA, Akihiro
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ELLA
exopolysaccharide
lectin
polysaccharide
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container_end_page 173
container_issue 4.5
container_start_page 167
container_title BUNSEKI KAGAKU
container_volume 72
creator OSHIMA, Toshifumi
NAKAYAMA, Kai
TAKABAYASHI, Kumiko
IWAKI, Misato
UBE, Nana
NAKAJIMA, Akihiro
description 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.
doi_str_mv 10.2116/bunsekikagaku.72.167
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subjects ELLA
exopolysaccharide
lectin
polysaccharide
title Development of a New Quantitative Method for Polysaccharide by Sandwich Enzyme-Linked Lectin Assay Using Single Lectin Species
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