Protein Adsorption Loss─The Bottleneck of Single-Cell Proteomics

Area of Science:

• Proteomics
• Cellular Biology
• Biochemistry

Background:

• Single-cell proteomics offers deep molecular insights into cellular functions.
• Protein adsorption loss (PAL) is a major technical hurdle in single-cell proteomics.
• Current PAL mitigation strategies lack specificity and quantitative evaluation.

Purpose of the Study:

• To address the challenge of protein adsorption loss (PAL) in single-cell proteomics.
• To quantitate the impact of PAL on single-cell proteomics.
• To propose methods for benchmarking and validating PAL.

Main Methods:

• Analysis of existing knowledge on PAL, protein expression, and surface area in single-cell proteomics.
• Summary of current PAL solutions and methods for characterizing protein adsorption.
• Review of strategies from single-cell genomics for addressing measurement errors.

Main Results:

• PAL profoundly and selectively affects single-cell proteomics, though not previously quantified.
• Existing mitigation strategies are generic and lack specific efficacy evaluation for individual proteins.
• There is an urgent need to benchmark PAL at the proteome scale with single-protein resolution.

Conclusions:

• Quantifying and benchmarking PAL is crucial for advancing single-cell proteomics.
• Orthogonal single-cell proteomic techniques are proposed for PAL cross-validation.
• Addressing PAL will accelerate the development and application of single-cell proteomics.