Investigation of Peptides for Molecular Recognition of C-Reactive Protein–Theoretical and Experimental Studies

Abstract

We investigate the interactions between C-reactive protein (CRP) and new CRP-binding peptide materials using experimental (biological and physicochemical) methods with the support of theoretical simulations (computational modeling analysis). Three specific CRP-binding peptides (P2, P3, and P9) derived from an M13 bacteriophage have been identified using phage-display technology. The binding efficiency of the peptides exposed on phages toward the CRP protein was demonstrated via biological methods. Fibers of the selected phages/peptides interact differently due to different compositions of amino acid sequences on the exposed peptides, which was confirmed by transmission electron microscopy. Numerical and experimental studies consistently showed that the P3 peptide is the best CRP binder. A combination of theoretical and experimental methods demonstrates that identifying the best binder can be performed simply, cheaply, and fast. Such an approach has not been reported previously for peptide screening and demonstrates a new trend in science where calculations can replace or support laborious experimental techniques. Finally, the best CRP binderthe P3 peptidewas used for CRP recognition on silicate-modified indium tin oxide-coated glass electrodes. The obtained electrodes exhibit a wide range of operation (1.0–100 μg mL–1) with a detection limit (LOD = 3σ/S) of 0.34 μg mL–1. Moreover, the dissociation constant K d of 4.2 ± 0.144 μg mL–1 (35 ± 1.2 nM) was evaluated from the change in the current. The selectivity of the obtained electrode was demonstrated in the presence of three interfering proteins. These results prove that the presented P3 peptide is a potential candidate as a receptor for CRP, which can replace specific antibodies.

Challenges and Advances in Biomarker Detection for Rapid and Accurate Sepsis Diagnosis: An Electrochemical Approach

“…This method gains a faradic current and eliminates the non-faradic or charging current. DPV was used as a method for the detection of CRP involving the silanization of triethoxysilylpropyl succinic anhydride on the surface of an indium tin oxide electrode and in the use of a new anhydride terminal group to react with the antibody’s amine group [ 92 ]. The detection was based on the redox signal of a ferrocenodimethanol redox couple, and its intensity changes were monitored through DPV measurement.…”

A Point-of-Care Testing Device Utilizing Graphene-Enhanced Fiber Optic SPR Sensor for Real-Time Detection of Infectious Pathogens

“…where C is the concentration of the tested target pathogen; λ is the response wavelength corresponding to C; and λ 0 is the wavelength corresponding to blank samples without analytes. The limit of detection (LOD) is calculated as follows [33,34]:…”

Rapid Microfluidic Immuno-Biosensor Detection System for the Point-of-Care Determination of High-Sensitivity Urinary C-Reactive Protein

“…The most commonly used techniques for CRP detection in clinical laboratories include enzyme-linked immunosorbent assays (ELISAs) [ 6 , 7 , 8 ], immunoturbidimetric assays [ 9 , 10 , 11 ], latex immunoagglutination assays [ 12 , 13 ], fluorescence assays [ 14 , 15 , 16 ], chemiluminescence assays [ 17 , 18 ], surface plasmon resonance-based assays [ 19 ], and electrochemical assays [ 20 , 21 , 22 , 23 , 24 , 25 ]. However, these methods are time-consuming and require skilled operators.…”