A new visual screening assay for catalytic antibodies with retro-aldol retro-Michael activity.
Fast and convenient methods are required for the detection of novel catalysts. We have developed a new assay to allow direct visualization of retro-aldol retro-Michael catalytic activity and have demonstrated it with catalytic antibody 38C2. The assay is based on a catalytic cleavage of a physiologically stable substrate to release 3,4-cyclohexeneoesculetin. The latter then reacts with iron(III) to generate a non-soluble complex that precipitates in the form of a black dye. This assay may be used for screening new catalysts for retro-aldol retro-Michael activity with improved efficiency for specific prodrug activation.
Shamis
Marina
M
Department of Organic Chemistry, School of Chemistry, Raymond and Beverly Sackler Faculty of Exact Sciences, Tel-Aviv University, Tel Aviv 69978, Israel.
Barbas
Carlos F
CF
Shabat
Doron
D
D017151:Antibodies, Catalytic / Q000032:analysis*; D002384:Catalysis; D011232:Chemical Precipitation; D003116:Color; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D011355:Prodrugs / Q000138:chemical synthesis / Q000378:metabolism*; D013329:Structure-Activity Relationship
D017151:Antibodies, Catalytic; D011355:Prodrugs