Abstract:
Proteins, as crucial biological macromolecules, perform essential functions. The identification and quantification of protein biomarkers, as well as accurate measurement of target proteins in complex matrices, are vital for disease prevention, diagnosis, and treatment. With the ongoing development of life sciences, scientists have introduced several protein quantification methods, including ultraviolet-visible spectrophotometry, enzyme-linked immunosorbent assay, and mass spectrometry-based quantitative methods. However, for high molecular weight proteins, especially those exceeding 200 kDa, their complex structure and enormous molecular weight lead to complicated preprocessing or non-specific, incomplete reactions, causing inaccurate quantification results. Single particle counting quantifies by individually counting particles in a solution. This method offers higher sensitivity, better differentiation, and less interference than traditional methods. Furthermore, given its simple sample preparation and non-destructive nature of analysis, single particle counting is well-suited for quantifying high molecular weight proteins at the complete structural level. This paper provides a comprehensive summary of protein quantification methods, with a specific focus on three typical single particle counting techniques: single particle inductively coupled plasma mass spectrometry (SP-ICP-MS), single molecule array (Simoa), and electrospray differential mobility analysis-condensation particle counter (ES-DMA-CPC). The operating principles of these methods and their applications in protein quantification are systematically explained, and the paper concludes with a projection of future developments in the field of protein quantification based on single particle counting.