Chilewski Shannon D
Bristol Myers Squibb, Precision Medicine, Bioanalytical, and Translational Sciences, Princeton, NJ, USA.
Bioanalysis. 2025 Jun;17(11):759-765. doi: 10.1080/17576180.2025.2517530. Epub 2025 Jun 12.
Accurate measurement of protein concentration is crucial in biological research, where protein-based reagents play a key role in assay performance and reliability. Traditional methods of protein quantification often measure total protein concentration, failing to account for the active portion capable of binding to its intended target. Calibration-free concentration analysis (CFCA), which uses surface plasmon resonance (SPR) technology, offers a solution by specifically measuring the active protein concentration of the sample. CFCA leverages binding under partially mass-transport limited conditions to directly quantify the functional protein in a sample, overcoming variability associated with recombinant protein production. This article provides a background on CFCA and the case for its more widespread use in protein reagent characterization, as it offers a way to reduce lot-to-lot and vendor-to-vendor variability while improving reproducibility and standardization of assays. This perspective was informed by searching and selecting pertinent articles from PubMed (Nov 2024-March 2025) and by examining the reference lists of key papers.
在生物学研究中，准确测量蛋白质浓度至关重要，因为基于蛋白质的试剂在检测性能和可靠性方面起着关键作用。传统的蛋白质定量方法通常测量总蛋白质浓度，而没有考虑到能够与其预期靶点结合的活性部分。无校准浓度分析（CFCA）利用表面等离子体共振（SPR）技术，通过专门测量样品中的活性蛋白质浓度提供了一种解决方案。CFCA利用部分传质受限条件下的结合来直接定量样品中的功能性蛋白质，克服了与重组蛋白质生产相关的变异性。本文提供了CFCA的背景信息以及在蛋白质试剂表征中更广泛使用它的理由，因为它提供了一种减少批次间和供应商间变异性的方法，同时提高了检测的重现性和标准化。这一观点是通过从PubMed（2024年11月至2025年3月）搜索和选择相关文章以及查阅关键论文的参考文献列表而形成的。
