Single-molecule force spectroscopy: A facile technique for studying the interactions between biomolecules and materials interfaces

Abstract

The quantification of the interactions between biomolecules and materials interfaces is crucial for design and synthesis functional hybrid bionanomaterials for materials science, nanotechnology, biosensor, biomedicine, tissue engineering, and other applications. Atomic force spectroscopy (AFM)-based single-molecule force spectroscopy (SMFS) provides a direct way for measuring the binding and unbinding forces between various biomolecules (such as DNA, protein, peptide, antibody, antigen, and others) and different materials interfaces. Therefore, in this review, we summarize the advance of SMFS technique for studying the interactions between biomolecules and materials interfaces. To achieve this aim, firstly we introduce the methods for the functionalization of AFM tip and the preparation of functional materials interfaces, as well as typical operation modes of SMFS including dynamic force spectroscopy, force mapping, and force clamping. Then, typical cases of SMFS for studying the interactions of various biomolecules with materials interfaces are presented in detail. In addition, potential applications of the SMFS-based determination of the biomolecule-materials interactions for biosensors, DNA based mis-match, and calculation of binding free energies are also demonstrated and discussed. We believe this work will provide preliminary but important information for readers to understand the principles of SMFS experiments, and at the same time, inspire the utilization of SMFS technique for studying the intermolecular, intramolecular, and molecule-material interactions, which will be valuable to promote the reasonable design of biomolecule-based hybrid nanomaterials.

Molecular Recognition of Surface Trans-Sialidases in Extracellular Vesicles of the Parasite Trypanosoma cruzi Using Atomic Force Microscopy (AFM)

“…In the case of EVs, AFM represents an attractive alternative for the determination of the morphology, structure and composition, and for the quantification of biophysical characteristics (stiffness, Young's modulus, and adhesion force, work of adhesion, hys-teresis, dissipation, and relaxation times). In particular, AFM-based single molecule-force spectroscopy is the only technique for detecting the molecular recognition forces between two molecules with subnanometric precision and high sensitivity of the order of nanometers and picometers [25][26][27][28]. This technique requires the functionalization of the tip of the cantilever with a molecule (ligand, receptor or antibody) that will approach until contact with a surface in which the molecule to which they bind and interact (receptor, ligand or antigen) is immobilized [29][30][31].…”

Molecular Crowding: The History and Development of a Scientific Paradigm

“…Whereas deuteration in principle allows to amplify the scattering contrast of a single component, variation of the solvent composition by changing the ratio of deuterated and hydrogenated solvent molecules may lead to full contrast matching of all but one component, thus enabling scientists to address the morphology of a single component in the mixture. The latter technique had been successfully applied to investigate the structure of macromolecules in a crowded (but invisible) environment. − Techniques such as single-molecule force spectroscopy, fluorescence-based assays, and advanced imaging methods can also provide valuable insights into the kinetics, thermodynamics, and spatial organization of intermolecular interactions in crowded environments. − However, challenges exist in accurately interpreting the experimental data due to the complexity of the crowded environment and the multitude of factors influencing it.…”

Development of Single Molecule Techniques for Sensing and Manipulation of CRISPR and Polymerase Enzymes

“…[ 107 ] To measure the interaction forces, the surface of an AFM cantilever tip is modified and functionalized with specific biomolecules using different chemical linkers (such as N‐hydroxysuccimidyl (NHS)‐PEG‐NHS, NHS‐PEG‐maleimide, thiol‐PEG‐NHS, or acetal‐PEG‐NHS). [ 108 ] A different secondary biomolecule is then tethered onto the surface. Figure a shows two such functionalization examples.…”