Shida Hiroaki, Naito Takafumi, Shibata Kaito, Yamada Yasuhide, Kawakami Junichi
Department of Hospital Pharmacy, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan.
Department of Clinical Oncology, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka, Japan.
Bioanalysis. 2018 Sep 1;10(18):1501-1510. doi: 10.4155/bio-2018-0161. Epub 2018 Sep 10.
Proteomics-based LC-MS/MS methods using trypsin solution have some problems including ion suppression and long protein digestion times. Few practical methods to quantify denosumab in human serum have been published.
Immunoglobulins in serum were extracted using immobilized protein G. Denatured, reduced and alkylated serum samples were digested with immobilized trypsin for 14 min. A denosumab-unique peptide was identified using a Fourier transform mass spectrometer as a signature peptide. The signature peptide was quantitated with a hybrid triple-quadrupole/linear ion-trap mass spectrometer.
A rapid and practical proteomics-based LC-MS/MS method using immobilized trypsin for denosumab quantitation in human serum was developed. The present method has an acceptable analytical performance and can be helpful for the determination of serum denosumab in clinical settings.
使用胰蛋白酶溶液的基于蛋白质组学的液相色谱-串联质谱（LC-MS/MS）方法存在一些问题，包括离子抑制和较长的蛋白质消化时间。目前已发表的用于定量人血清中地诺单抗的实用方法很少。
使用固定化蛋白G提取血清中的免疫球蛋白。将变性、还原和烷基化的血清样品用固定化胰蛋白酶消化14分钟。使用傅里叶变换质谱仪鉴定地诺单抗独特肽作为特征肽。用混合三重四极杆/线性离子阱质谱仪对特征肽进行定量。
建立了一种基于蛋白质组学的快速实用的LC-MS/MS方法，使用固定化胰蛋白酶定量人血清中的地诺单抗。本方法具有可接受的分析性能，有助于临床环境中血清地诺单抗的测定。
