Exploration of Semiconductor Chip-Based Single-Molecule Protein Sequencing for Identification of Hemoglobin Variants

Identification of hemoglobin (Hb) variants is of significant value in the clinical diagnosis of hemoglobinopathies. Conventional methods used to identify Hb variants in clinical laboratories can narrow down the range of candidates for a Hb variant sample but are unable to pinpoint the exact Hb variant. In this study, Next-Generation Protein Sequencing (NGPS), a semiconductor chip-based single-molecule protein sequencing (SMPS) technology, was explored as a novel method to identify Hb variants. Two heterozygous Hb variant samples underwent NGPS analysis. Proteotypic peptides corresponding to Hb variants were successfully detected, enabling the identification of the samples as Hb Handsworth (Hb α-Handsworth subunit G18R) and Hb G-Accra (Hb β-G-Accra subunit D73N). The NGPS method has been demonstrated as a potential tool to identify Hb variants. Although there are still limitations to overcome for the wide adoption of NGPS, this exploration supports the potential use of NGPS and other SMPS technologies in clinical applications.