Filter spot-ELISA for the enumeration of interferon-alpha antibody-secreting cells.
A filter spot-ELISA was developed for the enumeration of interferon-alpha (IFN-alpha) antibody secretion by murine and human cells. Various human IFN-alpha subtypes were coated onto nitrocellulose membranes by means of broad specificity bovine antibodies. Membranes were blocked with milk proteins, and antibodies released by individual cells during a 3 h culture period were visualized as distinct spots using peroxidase-conjugated, species-specific anti-immunoglobulin antibodies and diaminobenzidine/hydrogen peroxide substrate solution. The spot-ELISA is rapid, easy to perform, and highly sensitive. Possible applications include frequency estimates of IFN-alpha antibody-secreting cells in blood, tissues and hybridoma cultures as well as the evaluation of specificity and immunoglobulin class of the respective antibodies.
Prümmer
O
O
Department of Medicine III, University of Ulm, F.R.G.
Cederblad
B
B
Alm
G
G
Drees
N
N
Porzsolt
F
F
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000921:Antibody-Producing Cells / Q000276:immunology*; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D007370:Interferon Type I / Q000276:immunology*; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000276:immunology; D051379:Mice
D000911:Antibodies, Monoclonal; D007370:Interferon Type I