Squaric monoamide monoester as a new class of reactive immunization hapten for catalytic antibodies.
A squaric monoester monoamide motif was employed as an effective reactive immunogen for the discovery of monoclonal antibodies with reactive residue(s) in their combining sites. Two antibodies, 2D4 and 3C8, were uncovered that enhance paraoxon hydrolysis over background. Kinetic analysis of these antibodies was performed and interestingly both undergo a single turnover event due to covalent modification within the antibody combining site. Because antibodies 2D4 and 3C8 result in covalent attachment and thus inactivation of paraoxon, they could be useful probes for investigating paraoxon intoxication.
Xu
Yang
Y
The Skaggs Institute for Chemical Biology, Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Yamamoto
Noboru
N
Ruiz
Diana I
DI
Kubitz
Diane S
DS
Janda
Kim D
KD
D000577:Amides; D000818:Animals; D017151:Antibodies, Catalytic / Q000096:biosynthesis / Q000494:pharmacology*; D001666:Binding Sites, Antibody; D003503:Cyclobutanes / Q000008:administration & dosage / Q000138:chemical synthesis / Q000276:immunology*; D004952:Esters; D006241:Haptens / Q000008:administration & dosage / Q000737:chemistry* / Q000276:immunology; D006868:Hydrolysis; D007700:Kinetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010261:Paraoxon / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000276:immunology*; D010575:Pesticides / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000276:immunology; D013329:Structure-Activity Relationship; D014611:Vaccination
D000577:Amides; D017151:Antibodies, Catalytic; D003503:Cyclobutanes; D004952:Esters; D006241:Haptens; D010575:Pesticides; D010261:Paraoxon; C030042:squaric acid