Yong Xie

Proteoforms are proteins derived from highly related genes or post translational modifications (PTMs) of the same protein. They share extremely similar primary structures but have varying functions. Unfortunately, protein de novo sequencing including specific PTM/mutation detection is still challenging. Herein, a nanopore‐based technique is reported to resolve the amino acid order of amyloid‐β (Aβ_1‐42) with site specificity. Subnanopores are sputtered in 5 nm‐thick inorganic membranes with a sensing depth of 0.66 nm inferred by finite element analysis. Denatured molecules at 0.45 ng mL⁻¹ translocate through subnanopores while the current traces are sampled at 500 kHz with rms noise 1‐42 at single residue resolution, with Pearson Correlation Coefficients (PCCs) of 0.81 ± 0.03 and 0.92 ± 0.03 before and after dynamic time warping (DTW). A scrambled version of Aβ_1‐42 is tested for validation purposes. Deep learning classification reveals that different polypeptides generate distinct translocation fluctuating patterns, but variations become imperceptible for the same species measured across nanopores (Area Under the Curve, AUC 0.93 ± 0.05 vs 0.64 ± 0.12). Lastly, important PTMs and mutations are site‐specifically located along the primary structure, implying new potential clinical applications.

This paper reports a handy technical scheme to decorate atomic force microscopy (AFM) tips toward tip-enhanced Raman spectroscopy (TERS) applications. The major attraction of these homemade tips lies in that silver decoration can be confined at the apex of commercial tips by the means of an AFM-controlled electrochemical reaction. The reduction of Ag⁺ occurs in a highly sealed environment to secure the metal coating efficiency. Key factors include silver nitrate solution to provide Ag⁺, ambient relative humidity and temperature in a humidity cell, electric potential bias, and tip-surface distance. Subsequently, these silver-coated tips are evaluated for TERS measurement of carbon nanotubes (CNTs) so that both morphological and chemical characteristics of CNTs are concurrently obtained. The Raman spectra reveal that our plasmonic tip competently possesses an ∼30-fold local field signal increase and the corresponding TERS image laterally resolves at the single-pixel level.