Array-Based Rational Design of Short Peptide Probe-Derived from an Anti-TNT Monoclonal Antibody

Abstract

Complementarity-determining regions (CDRs) are sites on the variable chains of antibodies responsible for binding to specific antigens. In this study, a short peptide probe for recognition of 2,4,6-trinitrotoluene (TNT), was identified by testing sequences derived from the CDRs of an anti-TNT monoclonal antibody. The major TNT-binding site in this antibody was identified in the heavy chain CDR3 by antigen docking simulation and confirmed by an immunoassay using a spot-synthesis based peptide array comprising amino acid sequences of six CDRs in the variable region. A peptide derived from heavy chain CDR3 (RGYSSFIYWF) bound to TNT with a dissociation constant of 1.3 μM measured by surface plasmon resonance. Substitution of selected amino acids with basic residues increased TNT binding while substitution with acidic amino acids decreased affinity, an isoleucine to arginine change showed the greatest improvement of 1.8-fold. The ability to create simple peptide binders of volatile organic compounds from sequence information provided by the immune system in the creation of an immune response will be beneficial for sensor developments in the future.

Peptide-modified Microelectrode-based Potentiometric Device for 2,4,6-trinitrotoluene Molecule Detection

“…In accordance with a previous study, the 15-mer amino acid sequence (ARGYSSFIYWFFDFC) obtained from the complementarity-determining region heavy chain (HCDR) in the anti-TNT monoclonal antibody was determined as the TNT recognition peptide (named TNTHCDR3), whereas the TNTLCDR3 (sequence: CLQHYSAPYTC) peptide derived from the light chain of the complementarity-determining region (LCDR) in the anti-TNT monoclonal antibody was demonstrated to show no appreciable binding to the TNT explosive. (18) 1000 ppm TNT recognition peptide was immobilized through the amine coupling method to obtain the highest surface density for TNT binding, which was followed by blocking with ethanolamine solution. The surface morphology with/without the anchored peptide was evaluated by atomic force microscopy (AFM) as shown in Fig.…”

“…The TNT recognition peptide was synthesized in accordance with our previous work. (18,19) The mechanism for TNT explosive detection is based on the change in potential, which is mainly caused by the charge transfer between the positively charged group in TNT molecules and the negatively charged TNTHCDR3 peptide during the binding interaction. The sensitivity, selectivity, and stability of the peptide-based potentiometric device were investigated to evaluate its performance.…”

Enhanced Tubulation of Liposome Containing Cardiolipin by MamY Protein from Magnetotactic Bacteria

“…Peptide library of 15‐amino‐acid peptides was constructed by overlapping two amino acids along the sequence of MamY protein and synthesized on cellulose membranes (grade 542; Whatman, Maidstone, UK) activated with β‐alanine as N‐terminus by peptide auto‐spotter (MultiPep Rsi, Intavis AG, Köln, Germany) as shown in previous works . It should be noted that the peptide array comprising the 15‐mer peptides could reveal the importance of native secondary protein structure .…”

Bio-Inspired Strategies for Improving the Selectivity and Sensitivity of Artificial Noses: A Review

“…First, several teams have been trying to extract the sequences responsible for VOC recognition in ORs [49][50][51][52][53], OBPs [54][55][56][57], or antibodies [58]. As an example, presented in Figure 3, authors [57] used docking simulations on an OBP to extract peptides selective for octanal, and tested the influence of the chain length on the selectivity.…”