研究目的
To characterize and differentiate proteins with different shapes (spherical con.A and prolate Bovine Serum Albumin) at single-molecule level using solid-state nanopore technology.
研究成果
The study successfully differentiated between spherical and non-spherical proteins using solid-state nanopore technology. The method has potential applications in fingerprinting single proteins, offering a low-cost and high-throughput approach for protein characterization.
研究不足
The study is limited by the precision of nanopore detection and the conditions under which the experiments were conducted, such as the applied bias voltage and the ionic strength of the solution. Future improvements in detection precision could provide more accurate observations of protein dynamics.
1：Experimental Design and Method Selection:
The study utilized solid-state nanopore technology to investigate the translocation of proteins under an electric field. The methodology involved analyzing ionic current blockades caused by proteins passing through the nanopore.
2：Sample Selection and Data Sources:
Sphere-like con.A and prolate Bovine Serum Albumin (BSA) were selected as samples. The proteins were prepared in 1 M KCl and 10 mM phosphate buffered saline (PBS) solutions.
3：List of Experimental Equipment and Materials:
A patch-clamp amplifier (EPC 10 USB, HEKA Instruments) was used to apply constant voltage and acquire ionic current signals. A dual beam microscope (Helios 600i NanoLab, FEI Co.) was used for nanopore fabrication.
4：Experimental Procedures and Operational Workflow:
Proteins were electrophoretically driven through the nanopore under applied voltage, and ionic current signals were recorded. Molecular dynamics simulations were conducted to understand protein dynamics inside the nanopore.
5：Data Analysis Methods:
The ionic current blockade levels were analyzed to differentiate between spherical and non-spherical proteins. Gaussian distribution was used to fit the histogram of relative current blockade.
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