Development and Validation of an ELISA Method for Quantification of HuB01 in Fermentation Products

BACKGROUND: The antibodies (including 7C4, 8D1) are neutralizing antibodies specifically targeting the protein RBD, a key core of the combination with angiotensin-converting enzyme 2 (ACE2). In order to measure HuB01 in fermentation products accurately, a bioanalytical method was needed, which should have high selectivity and be free from interference by 501Y.V2. METHODS AND RESULTS: To quantify HuB01 in fermentation products, an indirect ELISA was developed and validated, utilizing anti-idiotypic monoclonal antibodies for both capture and detection. The assay demonstrates high sensitivity, with a lower limit of quantification set at 3.906 ng/mL, a wide dynamic range spanning 3906-125 ng/mL for HuB01, and outstanding precision and accuracy. CONCLUSION: A sensitive and selective method for measuring HuB01 in fermentation products has been developed and validated by us. Currently, this assay is applied in the quality control of the production process.