Drug-Protein Interactions for Clinical Research by Nucleic Acid Programmable Protein Arrays-Quartz Crystal Microbalance with Dissipation Factor Monitoring Nanoconductometric Assay

Abstract

Conductometric monitoring of drug-gene and drug-protein interactions is of fundamental importance in the field of molecular pharmacology. Here, we present our main findings and characterizations of an important antiblastic used in neuro-oncology (Temozolomide), interacting with selected proteins that represent predictive biomarkers of the rate survival of the patients, of the outcome of chemotherapy and resistance to drug itself (namely, BRIP1 and MLH1). We use our previously introduced two genes along with previously described Nucleic Acid Programmable Protein Arrays (NAPPA)-based nanoconductometric sensor. We performed a positive control (Temozolomide plus MLH1 protein), a negative control (Temozolomide plus BRIP1 protein) and a multi-gene experiment (Temozolomide plus BRIP1&MLH1 being coexpressed), showing that we are able to properly perform pharmacoproteomics tasks, discriminating each protein and drug unique conductance curve as well as their interactions, even in the presence of multi-proteins being immobilized. Moreover, in the last part of our paper, we used a multiple regression model in order to predict the behavior of Temozolomide when exposed to BRIP1&MLH1 co-expressed and we showed that we are able to predict the drug-protein interaction profile with a good regression coefficient.

Determination of Protein-Protein Interaction for Cancer Control via Mass Spectrometry and Nanoconductimetry of NAPPA SNAP Arrays: An Overview

“…We, thus, demonstrated the versatility of the NAPPA-QC biosensors for the detection of protein-protein interactions, protein-sterol and protein-drug interaction. Due to the simplicity with which new NAPPA-QC biosensors can be generated and to the successful application on clinical research [39], we envision the use of this platform for the development of biosensors for other applications, including, but not limited to, protein-small molecules, protein-lipids and protein-DNA.…”

NAPPA as a Real New Method for Protein Microarray Generation

“…Their results shower a properly discrimination of protein-protein interactions depending on the behavior against TMZ [15]. Finally, Liang et al .…”

“…Nicolini and collaborators clinically screened neuro-oncological patients respondent to temozolomide (TMZ) from those showing resistance to the drug by using a NAPPA-based nanoconductometric sensor [ 14 ]. Their results shower a properly discrimination of protein-protein interactions depending on the behavior against TMZ [ 15 ]. Finally, Liang et al .…”

Use of Protein Thin Film Organized by External Electric Field as a Template for Protein Crystallization

“…Another way of promoting nucleation and crystal growth is described in works with protein thin films. − In these studies, protein thin films are formed by the Langmuir–Blodgett (LB) technique, in which the protein solution is applied in a water–air interface followed by immediate submission to high pressure and then transferred to a glass slide. On this film, a small drop of protein solution is applied and the slide is reversed on the crystallization plate, as in the hanging drop vapor-diffusion method.…”