Analysis of active antibody concentration. Separation of affinity and concentration parameters.
Antibody binding to surfaces with differing amounts of immobilised antigen was measured in a biosensor system using surface plasmon resonance detection. Binding rates obtained during the initial binding phase on high density antigen surfaces were proportional to antibody concentration and independent of antigen-antibody affinity. One antibody calibration curve covering the range from 0.5 to 160 nM (0.08-25 micrograms/ml) antibody was valid for IgG antibodies with different antigen specificities. To illustrate the use of this methodology active antibody concentrations were analysed in culture media and in rabbit serum.
Karlsson
R
R
Pharmacia Biosensor AB, Uppsala, Sweden.
Fägerstam
L
L
Nilshans
H
H
Persson
B
B
D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000097:blood; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D000937:Antigen-Antibody Reactions; D015374:Biosensing Techniques; D005069:Evaluation Studies as Topic; D007118:Immunoassay / Q000379:methods*; D007700:Kinetics; D011817:Rabbits; D013806:Theophylline / Q000276:immunology
D000906:Antibodies; D000911:Antibodies, Monoclonal; D013806:Theophylline