The multiple antigen blot assay (MABA): a simple immunoenzymatic technique for simultaneous screening of multiple antigens

We describe a simple, reliable, reproducible and inexpensive technique termed multiple antigen blot assay (MABA) that permits the simultaneous screening of 28 different antigens based on a dot-blot ELISA methodology. Using an acrylic device (Miniblotter) containing 28 parallel troughs, multiple antigens are distributed and immobilized onto a nitrocellulose membrane. Strips are then cut perpendicularly and exposed to immune serum. The reaction is detected with secondary antibodies conjugated to horseradish peroxidase, and developed by a chemiluminescent substrate, the results being recorded on film. Positive reactions to the different antigens are seen as small black squares on each strip. We have used this qualitative technique to screen synthetic peptides derived from native schistosomal and malarial proteins, using immune rabbit sera as the detection antibody. This system can also be used in the clinical laboratory according to a dipstick-based diagnostic format for different infectious and non-infectious diseases, and can be designed to detect either antibodies or circulating antigens.