Catalytic polyclonal antibodies.
The production and characterization of rabbit polyclonal antibodies that catalyze the hydrolysis of trityl protecting groups is described. The catalytic activity increased throughout the immunization regimen, exhibiting a later and steeper rise than the simple hapten-specific antibody titer. The catalytic polyclonal antibodies displayed saturation behavior, consistent with classic Michaelis-Menton kinetics. An apparent catalytic rate enhancement (kcat/kuncat) of 125 and a Km of 31 microM were measured for the polyclonal sample. Inhibition studies indicated that 12% of the antibodies either binds the hapten or is a catalyst. Analyzing polyclonal antibodies for catalytic activity is proposed as a substantially faster, more cost effective and more general way to screen new hapten designs for their relative abilities to produce antibodies with catalytic activity.
Stephens
D B
DB
Department of Chemistry and Biochemistry, University of Texas, Austin 78712.
Iverson
B L
BL
D000818:Animals; D000906:Antibodies / Q000378:metabolism*; D002384:Catalysis; D006241:Haptens / Q000494:pharmacology; D007136:Immunoglobulins / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism*; D007202:Indicators and Reagents; D007700:Kinetics; D011817:Rabbits / Q000276:immunology
D000906:Antibodies; D006241:Haptens; D007136:Immunoglobulins; D007202:Indicators and Reagents