A high-throughput multi-species platform using Biolayer Interferometry Immunosorbent Assay (BLI-ISA) as an...

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. BLI-ISA provides rapid relative measurements of antigen-specific antibody levels (in nm binding shift), yielding results consistent with ELISA endpoint titers (expressed in ELISA Units), thereby enabling reliable comparison across treatment groups within a fraction of the time required for conventional ELISA.