Probe for selectively characterizing enzymes involved in xenobiotic metabolism and method of making and using the same

An assay platform, comprising:

• a substrate;
• and
• a probe having a structure satisfying Formula II
• wherein
• ERG, if present, is
• S(O)2OH, or an anionic form thereof;
• P(O(OH)2, or an anionic form thereof;
• a halogen;
• or —OPh-CH2—ONO2;
• EBG has a structure satisfying one or more of Formulas IIAEBG-IIJEBG
• wherein Y is CH3 or CF3;
• Y′ is O, NO2, or —N═NR″, wherein R″ is a dye or other reporting moiety;
• and m is an integer selected from 0 to 5;
• and R′ is selected from aldehyde, ketone, ester, carboxylic acid, acyl, acyl halide, cyano, sulfonate, nitro, nitroso, quaternary amine, CF3, alkyl halide, or combinations thereof;
• Linkera comprises an aliphatic group, a heteroaliphatic group, an aromatic group, an aliphatic-aromatic group, a heteroaliphatic-aromatic group, a heteroaromatic group, an aliphatic-heteroaromatic group, or a heteroaliphatic-heteroaromatic group;
• each of Linkerb, and Linker, if present, independently comprises an aliphatic group, a heteroaliphatic group, an aromatic group, an aliphatic-aromatic group, a heteroaliphatic-aromatic group, a heteroaromatic group, an aliphatic-heteroaromatic group, or a heteroaliphatic-heteroaromatic group;
• R, if present, is a group comprising a carbamate or a carbonate;
• each Tag, if present, independently comprises a functional group or molecule capable of generating a detectable signal;
• each pTag, which is present if a Tag is not present, independently comprises a clickable functional group;
• and
• each of n, m, p, and q independently is 0 or 1;
• and
• wherein the probe is covalently bound to the substrate.