A rapid method of epitope analysis using Superose 12 gel filtration. A study with monoclonal antibodies to chicken riboflavin carrier protein.
A novel and rapid method is described for determining the epitope specificities of monoclonal antibodies. The method utilises a highly reproducible, wide range gel filtration medium, Superose 12. nmol of monoclonal antibodies are incubated with pmol quantities of radiolabelled antigen, and the mixture filtered through Superose 12. Fractions are collected and radioactivity monitored. The size of the resultant immune complex indicates whether two monoclonal antibodies (mixed with the antigen) recognise the same or different epitope(s) of the antigen. The applicability of the above analytical method is illustrated with monoclonal antibodies raised against chicken egg riboflavin carrier protein.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000032:analysis; D002352:Carrier Proteins / Q000276:immunology*; D002645:Chickens; D002850:Chromatography, Gel / Q000379:methods*; D000939:Epitopes; D026901:Membrane Transport Proteins; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002352:Carrier Proteins; D000939:Epitopes; D026901:Membrane Transport Proteins; C019079:riboflavin-binding protein