A High-throughput Multi-Species Platform Using Biolayer Interferometry Immunosorbent Assay (BLI-ISA) as an Alternative to Indirect ELISA for Vaccine Development

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Abstract In vaccine development, the ELISA (Enzyme-Linked Immunosorbent Assay) is commonly used to compare the antibody titers of samples from several treatment groups. This often requires extensive sample preparation, manual labor, and long incubation and processing times. Biolayer Interferometry (BLI) has emerged as an alternative to the ELISA for the detection and quantification of antigen-specific antibodies in biological samples. However, the implementation of BLI as a replacement for the ELISA in vaccine development requires that experimental parameters are established for accurate and reproducible results. Here we give a general protocol for a biolayer interferometry immunosorbent assay (BLI-ISA) for the comparison of antigen-specific antibody levels in treatment group sera that uses secondary antibody binding responses as replacement for ELISA endpoint titers. We also validate that this BLI-ISA yields the same results as the ELISA endpoint titer while requiring far less time and effort. Competing Interest Statement WLP and WDP are affiliated with/cofounders of Hafion, Inc. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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last seen: 2026-05-20T01:45:00.602351+00:00