A CBM-based ELISA for HA was established according to the ELISA schemes, utilizing a hyaluronic acid-specific CBM. After optimization, the CBM-based ELISA for HA exhibited a linear detection range of 10 - 100 μg/mL. Both the intra- and inter-assay coefficients of variation were below 15%, ensuring the reliability of the results. The recovery rate ranged from 96.26% to 98.22%, indicating the accuracy of the method. The specificity of the method to HA demonstrated the reliance of CBM-based ELISA methods on CBM binding specificity. The effectiveness of the method in analyzing real samples containing hyaluronic acid was confirmed. Furthermore, a highly significant correlation was observed between the CBM-based ELISA and carbazole assay (y = 0.9951 x + 0.137, r = 0.994). By integrating CBM with ELISA, this study presented a novel and easily implementable solution for the simple and specific quantification of polysaccharides

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