Highly active enzyme preparations immobilized via matrix-conjugated anti-Fc antibodies

A novel method for enzyme immobilization to achieve highly active enzyme preparations is described. The enzyme is first reacted in solution with a specific monoclonal antibody (mAb) which does not interfere with its enzymic activity and which possesses a high affinity towards the enzyme, The immunocomplex is then reacted with immobilized anti-Fc antibodies which are coupled to the matrix by methods that preserve maximum immunological activity. Horseradish peroxidase (HRP) immobilization on epoxy-activated beads (CB6200) was chosen as a model system for this approach. Coupling of HRP via its mAbs to anti-Fc antibodies immobilized yielded the best results, corresponding to binding of 325 ng of fully active enzyme per bead. Binding of the enzyme to anti-HRP mAb directly immobilized on the beads was considerably lower.