Design and syntheses of three haptens to generate catalytic antibodies that cleave amide bonds with nucleophilic catalysis

Natural Catalytic Antibodies – Abzymes

“…A large COOH-terminal subsequence, VIP (15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28), was bound by the Ab with high affinity (K I = 1.25 nM), suggesting that it is the Ab-binding epitope. VIP (22)(23)(24)(25)(26)(27)(28), a short subsequence distant from the scissile bond, inhibited the binding (K I = 242 ìM) and hydrolysis (K I = 260 ìM) of full-length VIP by this Abz in a competitive fashion.…”

“…A large COOH-terminal subsequence, VIP (15)(16)(17)(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28), was bound by the Ab with high affinity (K I = 1.25 nM), suggesting that it is the Ab-binding epitope. VIP (22)(23)(24)(25)(26)(27)(28), a short subsequence distant from the scissile bond, inhibited the binding (K I = 242 ìM) and hydrolysis (K I = 260 ìM) of full-length VIP by this Abz in a competitive fashion. The auto-Ab did not show detectable binding of short VIP subsequences that encompass the scissile bond, VIP (15)(16)(17)(18)(19)(20)(21), VIP (11)(12)(13)(14)(15)(16)(17), and VIP (13)(14)(15)(16)(17)(18)(19)(20).…”

“…Alternatively, modification can be induced directly on purified antibody via selective chemical modification by the direct introduction of catalytic groups into the Ab combining site. Some studies describing these approaches are reported in [25][26][27][28][29] and references therein. The employment of the approaches has demonstrated that the substrate specificity (and/or the specific activity) of some artificial Abzs is comparable to or even higher than that of enzymes with the same catalytic activity [12,30,31].…”

A Catalytic Antibody Produces Fluorescent Tracers of Gap Junction Communication in Living Cells

“…Catalytic antibodies have proven utility for chemical catalysis in vitro (1)(2)(3)(4)(5)(6)(7). In living cells, these protein catalysts could accomplish a variety of reactions to modify cell behavior or generate reporter molecules for biological assays.…”

Disulfide Symmetric Dimers as Stable Pre‐Hapten Forms for Bioconjugation: A Strategy to Prepare Immunoreagents for the Detection of Sulfophenyl Carboxylate Residues in Environmental Samples

“…Instead, heterologous immunization with two individual haptens, containing a different charge, provides the opportunity to simultaneously generate acidic and basic catalytic residues in the antibody-combining site. Thus, Ersoy et al [46,47] designed three haptens to produce by heterelogous immunization nucleophile-mediated (phenol) amide bond-cleaving catalytic antibodies with a binding pocket with 1) a hydrophobic area, 2) an acidic residue complementary to the oxyanionic transition state, and 3) a basic residue to aid deprotonation of a phenol nucleophile and protonation of the departing amine. These examples point to the possibility of modulating the selectivity and affinity of the immunoresponse by combining immunoconjugates prepared by using two or more haptens without the need to invest time on complex synthetic procedures to preserve all the epitopes on the same hapten.…”