- 著者
- Sae SOTOMATSU Tomohiro YAMADA Hajime MIZUNO Hideki HAYASHI Toshimasa TOYO’OKA Kenichiro TODOROKI
- 出版者
- The Society for Chromatographic Sciences
- 雑誌
- CHROMATOGRAPHY (ISSN:13428284)
- 巻号頁・発行日
- pp.2019.013, (Released:2019-06-21)
- 参考文献数
- 16
- 被引用文献数
- 6
To construct liquid chromatography (LC)-based bioanalytical method for therapeutic monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs), twelve commercially available therapeutic mAbs and one ADC were chemically reduced, and the generated fragments were analyzed by high-temperature reversed-phase LC. For most therapeutic mAbs, single peaks of light and heavy chains were detected, indicating a possibility of homogeneous LC analysis using light chains. However, characteristic fragmentations were observed in infliximab, pembrolizumab, ramucirumab, and trastuzumab emtansine. We also performed a simple validation using the fragmented light chains for the bioanalysis of bevacizumab. The limit of detection (LOD) and limit of quantification (LOQ) of bevacizumab were 0.63 and 2.10 µg/mL, respectively, with dithiothreitol reduction, and 0.74 and 2.48 µg/mL, respectively, with tris (2-carboxyethyl) phosphine reduction. These results indicate that both the reductants confer sufficient linearity, LOQ, and LOD for the light chain analysis of bevacizumab. Thus, this method, combined with affinity purification, can be used for the bioanalysis of bevacizumab.
- 著者
- Kenichiro TODOROKI Yuto KUDOH Misuzu NAKAMURA Yuki SHIMIZU Takamitsu SASAKI Hideyuki OTSUKI Keisuke WADA Jun Zhe MIN Hajime MIZUNO Kouichi YOSHINARI Toshimasa TOYO’OKA
- 出版者
- The Japan Society for Analytical Chemistry
- 雑誌
- Analytical Sciences (ISSN:09106340)
- 巻号頁・発行日
- pp.18SCP05, (Released:2018-07-20)
- 被引用文献数
- 6
- 著者
- Kenichiro TODOROKI Tomohiro YAMADA Hajime MIZUNO Toshimasa TOYO’OKA
- 出版者
- The Japan Society for Analytical Chemistry
- 雑誌
- Analytical Sciences (ISSN:09106340)
- 巻号頁・発行日
- vol.34, no.4, pp.397-406, 2018-04-10 (Released:2018-04-10)
- 参考文献数
- 74
- 被引用文献数
- 26
The increase in the use of therapeutic monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) has made the detailed bioanalysis of these drugs essential not only for planning optimal therapeutic programs for clinical practice, but also for evaluating the biological equivalencies in the development of other biosimilars. The ligand binding assays that are widely in use now are being replaced rapidly by the highly accurate, sensitive, and selective analytical method using a mass spectrometer. This review will discuss the progress in and challenges observed during the development of a mass spectrometry-based bioanalytical method for therapeutic mAbs and ADCs.
- 著者
- Ryoko TOMITA Kenichiro TODOROKI Hiroshi MARUOKA Hideyuki YOSHIDA Toshihiro FUJIOKA Manabu NAKASHIMA Masatoshi YAMAGUCHI Hitoshi NOHTA
- 出版者
- The Japan Society for Analytical Chemistry
- 雑誌
- Analytical Sciences (ISSN:09106340)
- 巻号頁・発行日
- vol.32, no.8, pp.893-900, 2016-08-10 (Released:2016-08-10)
- 参考文献数
- 22
- 被引用文献数
- 1 12
We performed a comprehensive quantification of 20 amino acids in RPMI 1640 medium-cultured human colorectal adenocarcinoma cells to evaluate the efficacy of 5-fluorouracil treatment under hypoxic and hypoglycemic conditions, which mimic the tumor microenvironment. In this study, we developed a simple and comprehensive analytical method by using LC-MS/MS connected to the Intrada amino acid column, which eluted amino acids within 9 min. The present method covered a linearity range of 3.6 – 1818 μM, except for Gly (227 – 1818 μM), Ala, Asp, His (7.1 – 1818 μM each), and Trp (3.6 – 909 μM). The limits of detection were in the range of 0.02 – 38.0 pmol per injection in a standard solution. Amino acid concentration data were analyzed using principal-component analysis to represent samples on two-dimensional graphs. Linear discriminant analysis was used to classify samples on the score plots. Using this approach, the effect of 5-fluorouracil treatment could be successfully discriminated at high discrimination rates. Moreover, several amino acids were extracted from corresponding loading plots as candidate markers for distinguishing the effects of the 5-fluorouracil treatment or tumor microenvironmental conditions. These results suggest that our proposed method might be a useful tool for evaluating the efficacy of anticancer drugs in the tumor microenvironment.