Absolute protein quantification based on calibrated particle counting using electrospray-differential mobility analysis

化学 色谱法 肌红蛋白 粒径 粒子(生态学) 分析化学(期刊) 定量蛋白质组学 定量分析(化学) 生物系统 蛋白质组学 生物化学 海洋学 物理化学 地质学 生物 基因
作者
Wei Mi,Xinyi Zhang,Geyu Lu,Ruixue Sun,Shangying Ma,Zhishang Hu,Xinhua Dai
出处
期刊:Analytica Chimica Acta [Elsevier]
卷期号:1304: 342534-342534
标识
DOI:10.1016/j.aca.2024.342534
摘要

The traceability of in vitro diagnostics or drug products is based on the accurate quantification of proteins. In this study, we developed an absolute quantification approach for proteins. This method is based on calibrated particle counting using electrospray-differential mobility analysis (ES-DMA) coupled with a condensation particle counter (CPC). The absolute concentration of proteins was quantified with the observed protein particle number measured with ES-DMA-CPC, and the detection efficiency was determined by calibrators. The measurement performance and quantitative level were verified using two certificated reference materials, BSA and NIMCmAb. The linear regression fit for the detection efficiency values of three reference materials and one highly purified protein (myoglobin, BSA, NIMCmAb and fibrinogen) indicated that the detection efficiency and the particle size distribution of these proteins exhibited a linear relationship. Moreover, to explore the suitability of the detection efficiency-particle size curve for protein quantification, the concentrations of three typical proteinaceous particles, including two high molecular weight proteins (NIST reference material 8671 and D-dimer) and one protein complex (glutathione S-transferase dimer), were determined. This work suggests that this calibrated particle counting method is an efficient approach for nondestructive, rapid and accurate quantification of proteins, especially for measuring proteinaceous particles with tremendous size and without reference standards.
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