Department of Life Science and Chemistry, Jacobs University Bremen, Germany.
Department of Life Science and Chemistry, Jacobs University Bremen, Germany.
Mol Immunol. 2021 Aug;136:73-81. doi: 10.1016/j.molimm.2021.04.028. Epub 2021 Jun 3.
Recombinant major histocompatibility complex class I molecules are used in diagnostic and therapeutic approaches in cancer immunotherapy, with many studies exploring their binding to antigenic peptides. Current techniques for kinetic peptide binding studies are hampered by high sample consumption, low throughput, interference with protein stability, and/or high background signal. Here, we validate nanoscale differential scanning fluorimetry (nanoDSF), a method using the tryptophan fluorescence of class I molecules, for class I/peptide binding, and we use it to determine the molecular mechanism of the thermal denaturation of HLA-A*02:01.
重组主要组织相容性复合体 I 类分子被用于癌症免疫治疗中的诊断和治疗方法，许多研究都在探索它们与抗原肽的结合。目前用于研究动力学肽结合的技术受到高样品消耗、低通量、对蛋白质稳定性的干扰和/或高背景信号的限制。在这里，我们验证了纳米级差示扫描荧光法（nanoDSF），这是一种使用 I 类分子的色氨酸荧光的方法，用于 I 类/肽结合，并使用它来确定 HLA-A*02:01 热变性的分子机制。
