FLU-LISA (fluorescence-linked immunosorbent assay): high-throughput antibody profiling using antigen microarrays

Area of Science:

• Immunology
• Biotechnology
• Assay Development

Background:

• Humoral immune responses, crucial for protection against infections, are encoded by antibodies.
• Existing immunoassays often measure antibody responses to only a single antigen.
• Enzyme-linked immunosorbent assay (ELISA) is a common, semiquantitative method for antibody measurement.

Purpose of the Study:

• To introduce FLU-LISA, a novel antigen microarray-based assay for rapid, high-throughput antibody profiling.
• To enable simultaneous measurement of immunoglobulin (Ig) G, IgA, and IgM responses against multiple antigens.
• To provide a sensitive and specific alternative to traditional ELISA for antibody profiling.

Main Methods:

• Development of a fluorescence-linked immunosorbent assay (FLU-LISA) utilizing antigen microarrays.
• Profiling of IgG, IgA, and IgM responses to influenza and SARS-CoV-2 antigens.
• Comparison of FLU-LISA with traditional ELISA using samples from mice, chickens, and humans.
• Evaluation of FLU-LISA performance with dried blood spots.

Main Results:

• FLU-LISA demonstrated high specificity and sensitivity in profiling antibody responses.
• The assay successfully measured IgG, IgA, and IgM responses to multiple antigens simultaneously.
• FLU-LISA showed comparable or superior performance to ELISA across different sample types.
• The assay is compatible with dried blood spots, facilitating sample collection from humans and wild birds.

Conclusions:

• FLU-LISA is a novel, high-throughput assay for comprehensive antibody profiling.
• The assay offers a rapid and efficient alternative to traditional ELISA for various applications.
• FLU-LISA's compatibility with dried blood spots expands its utility in epidemiological studies and wildlife surveillance.