Immunizing Mice and Rats with Nitrocellulose-Bound Antigen

Abstract

A desired target antigen within a mixture of antigenic proteins such as a cell lysate can be separated from the mixture by gel electrophoresis. However, extracting the antigen from a section of gel and eluting it can be cumbersome and often results in significant loss of protein. Also, the animal might not tolerate the injection of the polyacrylamide gel piece containing the antigen. Alternatively, the protein can be transferred from the gel onto nitrocellulose and then injected either subcutaneously or into the peritoneal cavity. Protein antigens immobilized on nitrocellulose often make exceptionally good immunogens. This is likely because of their slow release from the paper, thus behaving somewhat like an adjuvant. Not all antigens show increased immunogenicity using this methodology, but some do. The antigen is bound to paper and can be implanted on the back of the mouse's neck, a location that makes it difficult for the mouse to disturb the surgical clip, or extracted and injected.