Mechanical characterization of HIV‐1 with a solid‐state nanopore sensor

Abstract

Enveloped viruses fuse with cells to transfer their genetic materials and infect the host cell. Fusion requires deformation of both viral and cellular membranes. Since the rigidity of viral membrane is a key factor in their infectivity, studying the rigidity of viral particles is of great significance in understating viral infection. In this paper, a nanopore is used as a single molecule sensor to characterize the deformation of pseudo-type human immunodeficiency virus type 1 at sub-micron scale. Non-infective immature viruses were found to be more rigid than infective mature viruses. In addition, the effects of cholesterol and membrane proteins on the mechanical properties of mature viruses were investigated by chemically modifying the membranes. Furthermore, the deformability of single virus particles was analyzed through a recapturing technique, where the same virus was analyzed twice. The findings demonstrate the ability of nanopore resistive pulse sensing to characterize the deformation of a single virus as opposed to average ensemble measurements.

Mechanical characterization of vesicles and cells: A review

“…In recent years, a lot of interest is being directed toward SSN for nanovesicle characterization since optical detection technique used in traditional electrodeformation studies fail to detect nanosized vesicles. Nanopore technology has been widely used since its inception nearly two decades ago for DNA , proteins , glycans , viruses , liposomes , nanoparticles , and polymer profiling because of its high resolution (single‐molecule level), low‐cost, high throughput, minimal sample requirement (both volume and concentration).…”

“…Recently, SSN assays were used to evaluate the electrodeformability of viruses and liposomes through a multiple recapture protocol . The multiple recapture protocol would evaluate the membrane mechanical properties of single‐vesicle multiple times (e.g., ∼25 times), thus, not only providing a statistically significant evaluation of the said properties of a single vesicle but also superseding conventional nanopore protocols which provide one value per translocating vesicle.…”

“…Atomic force microscopy (AFM) and other newer methods, such as optical and magnetic tweezers (MTs), have seen widespread use in characterizing this smaller breed of vesicles, although most of these techniques are low throughput . Electrodeformation assays with optical visualization is another popular tool in large vesicle characterization , while electrodeformation techniques with solid‐state nanopores (SSN) are now extending the limit of this useful nonintrusive method to nanosized vesicles .…”

Application of Solid-State Nanopore in Protein Detection

“…To date, in addition to promising applications in nucleic acids detection [36,[40][41][42][43][44][45], solid-state nanopores have made great progress in molecular interaction [46][47][48], detecting protein structures or their aggregation states [49][50][51][52], and virus identification [53]. However, nanopore signals of proteins are harder to resolve due to diversity of amino acids and inhomogeneous charge, as well as fast translocation [54].…”

Fabrication of hexagonal boron nitride based 2D nanopore sensor for the assessment of electro‐chemical responsiveness of human serum transferrin protein

“…Nanopores are ostensibly simple, label‐free, low‐cost overhead, and high‐throughput sensors used to profile a plethora of biomolecules and particles such as DNA , proteins , polysaccharides,, liposomes , and viruses with a high resolution. In its early days, just over two decades ago , and to a large extent even today, the biological counterpart dictated terms due to its size reproducibility, low noise, and vast genetic engineering protocols, despite limitations in available pore size and chemical and physical working conditions.…”