Medizinisches Proteom-Center (MPC), Medical Faculty, Ruhr-University Bochum, Bochum, Germany.
Medical Proteome Analysis, Center for Proteindiagnostics (PRODI), Ruhr-University Bochum, Bochum, Germany.
Methods Mol Biol. 2021;2228:21-28. doi: 10.1007/978-1-0716-1024-4_2.
Although a lot of new methods for protein concentration determination have been developed and established the last years, the amino acid analysis has still this relevance within proteomics for multiple reasons especially in the quantitative protein analysis. Amino acid analysis enables indirectly both the protein and peptide concentration determination which are essential for using the same amounts for comparative quantitative experiments. Moreover, the quantity and quality of synthetic peptides can be verified. The method itself is robust in comparison with colorimetric assays, especially when detergents or chaotropes are present in the sample buffer. Furthermore, it is highly sensitive. Nevertheless, amino acid analysis needs a certain experience to be set up and is time-consuming compared to other protein concentration determination techniques.
尽管近年来已经开发并建立了许多用于蛋白质浓度测定的新方法,但氨基酸分析在蛋白质组学中仍然具有重要意义,原因有很多,尤其是在定量蛋白质分析中。氨基酸分析可以间接测定蛋白质和肽的浓度,这对于使用相同量进行比较定量实验至关重要。此外,还可以验证合成肽的数量和质量。与比色测定法相比,该方法本身具有较强的稳健性,特别是在样品缓冲液中存在去污剂或变溶剂时。此外,它的灵敏度也很高。然而,与其他蛋白质浓度测定技术相比,氨基酸分析需要一定的经验才能建立,并且耗时较长。