Title : Selection of a human butyrylcholinesterase-like antibody single-chain variable fragment resistant to AChE inhibitors from a phage library expressed in - Podesta_2014_MAbs_6_ |
Author(s) : Podesta A , Rossi S , Massarelli I , Carpi S , Adinolfi B , Fogli S , Bianucci AM , Nieri P |
Ref : MAbs , 6 : , 2014 |
Abstract :
Organophosphates are potent poisoning agents that cause severe cholinergic toxicity. Current treatment has been reported to be unsatisfactory and novel antidotes are needed. In this study, we used a single-chain variable fragment (scFv) library to select a recombinant antibody fragment (WZ1-14.2.1) with butyrylcholinesterase-like catalytic activity by using an innovative method integrating genetic selection and the bait-and-switch strategy. Ellman assay demonstrated that WZ1-14.2.1 has Michaelis-Menten kinetics in the hydrolysis of all of the three substrates used, acetylthiocholine, propionylthiocholine and butyrylthiocholine. Notably, the catalytic activity was resistant to the following acetylcholinesterase inhibitors: neostigmine, iso-OMPA, chlorpyrifos oxon, dichlorvos, and paraoxon ethyl. Otherwise, the enzymatic activity of WZ1-14.2.1 was inhibited by the selective butyrylcholinesterase inhibitor, ethopropazine, and by the Ser-blocking agent phenylmethanesuphonyl fluoride. A hypothetical 3D structure of the WZ1-14.2.1 catalytic site, compatible with functional results, is proposed on the basis of a molecular modeling analysis. |
PubMedSearch : Podesta_2014_MAbs_6_ |
PubMedID: 24675419 |
Podesta A, Rossi S, Massarelli I, Carpi S, Adinolfi B, Fogli S, Bianucci AM, Nieri P (2014)
Selection of a human butyrylcholinesterase-like antibody single-chain variable fragment resistant to AChE inhibitors from a phage library expressed in
MAbs
6 :
Podesta A, Rossi S, Massarelli I, Carpi S, Adinolfi B, Fogli S, Bianucci AM, Nieri P (2014)
MAbs
6 :