Development of a potential primary method for protein quantification via electrospray differential mobility analysis.

Accurate protein quantification is the basis for establishing the metrological traceability of in vitro diagnostics or drug products. In this study, we established and validated a potential primary method for protein quantification based on electrospray-differential mobility analysis coupled with a condensation particle counter (ES-DMA-CPC). The analytical performance of this method was assessed using the certified reference material NIMCmAb, and the uncertainty of measurement was evaluated. The method was applied to the quantification of three other protein reference materials and one highly purified protein, including myoglobin, bovine serum albumin, IgG monoclonal antibody, and one highly purified fibrinogen, with a molecular weight range between 17 kDa and 340 kDa. In addition, when compared with isotope dilution mass spectrometry (IDMS) and UV‒VIS spectrophotometry approaches, the ES-DMA-CPC method showed good agreement with IDMS method for the quantification of these protein reference materials. Our proposed method provided an accurate quantification of proteins, especially those with large molecular weights. Moreover, our method could be a potential primary method for protein quantification and serve as a complement to IDMS method.