Simple ELISA using Polyclonal Antibody and Lectin for Determination of Soluble Chitosan - Like Polysaccharide in Bacterial CultureJournal: International Journal of BioAnalytical Methods & BioEquivalence Studies (IJBMBS) (Vol.04, No. 01)
Publication Date: 2017-12-07
Authors : Takeo M Taguchi S Odagaki S Baranwal P Kimura K Negoro S;
Page : 76-81
Keywords : ELISA; Chitosan; Biopolymer; Polysaccharides; Lectin; Citrobacter.;
We have developed a simple enzyme-linked immunosorbent assay (ELISA) for the direct determination of chitosan-like polysaccharide (CP) produced in the cultures of Citrobacter strains. CP in sample solutions was reacted with anti-CP rabbit polyclonal antibody immobilized in a 96-well microtiter plate and sandwiched with horseradish peroxidase-conjugated lectin, followed by color development by the enzyme activities. Using standard chitosan solutions, a good standard curve (R2=0.977) was established between 0 and 250 μg l-1 with the average coefficient of variation of 7.0%, and CP in the diluted culture supernatants could be directly determined. This method can apply for the direct determination of other polysaccharides in solution by selecting an inexpensive custom-made polyclonal antibody and an appropriate lectin, both of which may be commercially available.
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