An investigation of antibody acyl hydrolysis catalysis using a large set of related haptens.
An aspect of catalytic antibody research that receives little attention in the literature involves hapten systems that fail to elicit antibody catalysts despite a high affinity immune response and hapten designs that resemble those known to elicit catalysts. We have investigated a series of 12 phosphate and phosphonate haptens in a total of three animal systems. Dramatic and reproducible differences were observed in the catalytic activities of polyclonal antibodies elicited by the different haptens. A phosphate hapten with a phenyl ring on the side of the hapten opposite the linker elicited reproducibly high levels of polyclonal antibody catalytic activity. The other 11 haptens, most with benzyl groups on the side of the hapten opposite the linker, elicited immune responses in which catalytic activity was significantly weaker in terms of the level of observed catalytic activity, as well as frequency of elicited catalysts. Our results indicate that subtle features of transition state analogue hapten structure can have a dramatic and reproducible influence over the catalytic activity of elicited antibodies in related haptens. Whatever the explanation, subtle changes in mechanistic features due to altered leaving group ability/location or overall hapten flexibility, the comprehensive data presented here indicate that phenyl or 4-nitrophenyl leaving groups located opposite the hapten linker are to be preferred in order to elicit highly active antibody catalysts for acyl hydrolysis reactions.
Odenbaugh
A L
AL
The Department of Chemistry and Biochemistry, The University of Texas at Austin, 78712, USA.
Helms
E D
ED
Iverson
B L
BL
D000818:Animals; D017151:Antibodies, Catalytic / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D006241:Haptens / Q000378:metabolism*; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008807:Mice, Inbred BALB C; D008813:Mice, Inbred ICR; D016339:Spectrometry, Mass, Fast Atom Bombardment
D017151:Antibodies, Catalytic; D006241:Haptens