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

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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

Sachise KARAKAWA Masashi HARADA Rumi NISHIMOTO

出版者

The Society for Chromatographic Sciences

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

vol.44, no.1, pp.21-26, 2023-02-20 (Released:2023-03-11)

参考文献数

47

被引用文献数

3

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Three different types of analytical methods for D,L-amino acid were developed. First, a method for simultaneous analysis of D,L-amino acids in food was constructed by combining pre-column derivatization with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), UHPLC separation, and UV and circular dichroism (CD) detection. Using this method, D,L-amino acids in food samples was determined at a very short time of 5.5 minutes. Second, a pre-column derivatization LC/MS method (direct method) for D-alanine, D-serine, and D-aspartic acid and L-isomers in biological samples was developed. In this method, D,L-amino acids were derivatized with an achiral reagent, p-N,N,N-trimethylammonioanilyl N′-hydroxysuccinimidyl carbamate iodide (TAHS), separated with a chiral column, and detected using mass spectrometry (MS). Third, a pre-column derivatization LC/MS method (indirect method) for highly sensitive and selective analysis of 20 D,L-amino acids using a chiral derivatization reagent, (R)-4-nitrophenyl-N-[2-(diethylamino)-6,6-dimethyl-[1,1-biphenyl]-2-yl] carbamate hydrochloride ((R)-BiAC), was developed. The unique structure of the (R)-BiAC reagent, which has a biaryl axially chiral structure, allowed for higher resolution and a high sensitive analysis of the 20 D,L-amino acids within 12 minutes. This method has been applied to various D-amino acid research studies, and it can be applied to a wide range of analyses from food to biological samples. Lastly, an LC/MS method was established that simultaneously analyzes tryptophan and 15 of its metabolites, which is involved in neurotransmission and immunity in vivo.

著者

Naoki NISHIMURA Toyohiro NAITO Takuya KUBO Koji OTSUKA

出版者

The Society for Chromatographic Sciences

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

vol.39, no.3, pp.113-118, 2018-10-20 (Released:2018-11-08)

参考文献数

36

被引用文献数

6 4

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In this study, we reveal the suppression of non-specific hydrophobic interaction in poly(ethylene-co-glycidylmethacrylate) (PEGM) based spongy monolith (SPM), PEGM-SPM, which has recently be reported as a new platform of separation medium for affinity chromatography in our previous study, by an simple acidic treatment. Additionally, the immobilization procedures of protein-A toward the PEGM-SPM and the separation conditions for immunoglobulin G (IgG) were optimized for further effective affinity separations. As a result of treatment by a mixture of trifluoroacetic acid and acetonitrile, the hydrophobicity was dramatically suppressed in the PEGM-SPM. The optimizations for the density of PEGM in the PEGM-SPM, the protein A immobilization, and the binding/releasing conditions showed that variety of proteins and peptides were not retained on the protein A immobilized spongy column at all while IgG was absolutely separated by a simple stepwise pH gradient condition.

著者

Yasushi ISHIHAMA

出版者

The Society for Chromatographic Sciences

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

pp.2019.023, (Released:2019-10-10)

参考文献数

64

被引用文献数

10

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Mass spectrometry-based proteomics platforms have been widely used as ‘proteome sequencers’ to characterize the proteomes of a wide range of organisms. The work-flow generally involves multiple steps of sample preparation, peptide purification/concentration/pre-fractionation, and nanoLC/MS/MS measurement. This review focuses on our contributions to the technical development of current proteomics platforms, and includes a consideration of the limitations of these systems, together with the prospects for developing superior new-generation proteome sequencers.

著者

Takako HAYASHI Teruhisa FUJIMAKI Maki MIYAZAWA

出版者

The Society for Chromatographic Sciences

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

pp.2018.011, (Released:2018-09-29)

参考文献数

16

被引用文献数

4

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We examined the utility of LC-MS/MS/MS in the rapid quantitative analysis of clenbuterol in pig liver. Compared with LC-MS/MS in the SRM mode, which gave a chromatogram of pig liver containing interference peaks near the target component peak, LC-MS/MS/MS generated a clear chromatogram with no interference peaks. Validation of the method yielded favorable results for both LC-MS/MS in the SRM mode and LC-MS/MS/MS, with trueness values of 95.5% and 102%, repeatability of 1.9% and 5.5%, and within-laboratory reproducibility of 2.0% and 5.1%, respectively. Moreover, the recovery of clenbuterol-d9 used as surrogate was ≥70% in both measurement methods and thus, validation was achieved in terms of both selectivity and limit of quantification. The results suggest that LC-MS/MS/MS has highly selectivity for quantitative analysis as the influence of interference peaks is suppressed, and can be used when quantification is difficult with LC-MS/MS in the SRM mode due to matrix effects.

著者

Chiharu ISHII Tetsuya MIYAMOTO Shoto ISHIGO Yurika MIYOSHI Masashi MITA Hiroshi HOMMA Tadashi UEDA Kenji HAMASE

出版者

クロマトグラフィー科学会

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

pp.2017.009, (Released:2017-06-04)

参考文献数

46

被引用文献数

15

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The formation of D-amino acid residues in proteins is considered as one of the deterioration processes, and the determination of these D-amino acid residues is highly expected for the screening of new biomarkers under various disease conditions. In the present study, a two-dimensional (2D) HPLC-MS/MS system following the hydrolysis with deuterium chloride (2HCl/2H2O) and derivatization of amino acids with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) has been designed/developed and applied to the analysis of proteins stored under various conditions. As the target, 5 major D-amino acid residues (Ala, Asp, Glu, Pro and Ser) were selected. The analytical procedure was validated using a model peptide, NH2-Gly-Pro-Glu-Ala-Asp-Ser-Gly-OH, and the obtained calibration lines of %D for the 5 target amino acids were linear with correlation coefficients greater than 0.998. The RSD values for the intra-day precision and inter-day precision were lower than 5%. In most of the proteins tested, the amounts of the D-Ser and D-Asp residues increased during storage, and the highest value (14%, D-Ser) was observed in ovalbumin (OVA) after storage at pH 9.5 for 4 weeks.

著者

LIM Lee Wah

出版者

クロマトグラフィー科学会

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

pp.2015.011, (Released:2015-04-13)

参考文献数

77

被引用文献数

1 1

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This focusing review summarizes our recent works on the dissemination of capillary liquid chromatography through various approaches, i.e. simple stepwise-gradient elution system, on-line sample enrichment system, alternate-pumping recycle chromatography system, on-line immobilized-enzyme reactor, development of novel stationary phases, etc. Capillary liquid chromatography saves energy and waste, and it possesses several advantages over the conventional liquid chromatography; however, it has not been much popularized since its appearance and most users still prefer to use conventional liquid chromatography if there is a choice between them. Our research focuses on, but not limited to, developing versatile capillary liquid chromatography systems that are not only environmental-friendly but also user-friendly, and novel stationary phases that could be operated under mixed modes conditions by manipulating the eluent, which are applicable to real samples analyses will therefore eventually switch the users from “conventional” to “capillary”.

著者

Kenji HAMASE Yusuke NAKAUCHI Yurika MIYOSHI Reiko KOGA Nao KUSANO Hirohisa ONIGAHARA Hiroshi NARAOKA Hajime MITA Yasuhiko KADOTA Yasuhiro NISHIO Masashi MITA Wolfgang LINDNER

出版者

クロマトグラフィー科学会

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

vol.35, no.2, pp.103-110, 2014-08-10 (Released:2014-08-26)

参考文献数

39

被引用文献数

4 33

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A two-dimensional chiral high-performance liquid chromatographic (2D-HPLC) system has been established for the analysis of extraterrestrial amino acids. As the targets, 8 chiral amino acids (alanine (Ala), valine (Val), 2-aminobutyric acid (2AB), norvaline (nVal), N-methylalanine (N-MeAla), isovaline (iVal), 3AB and 3-aminoisobutyric acid (3AIB)) and 5 non-chiral amino acids (glycine (Gly), β-Ala, γ-aminobutyric acid (GABA), sarcosine (Sar) and 2AIB) were selected. These amino acids were tagged with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), and non-enantioselectively separated by a capillary monolithic ODS column in the first dimension. The target fractions were automatically introduced into the second dimension and further separated by Pirkle-type enantioselective columns. By using this system, the 2D-HPLC separation of 21 components in small particles of a carbonaceous chondrite (Yamato 791191, Antarctic CM2 meteorite) could be successfully performed, and all of the target amino acids were observed. The D/L ratios of the chiral molecules are almost 50/50 for all of the tested proteinogenic and non-proteinogenic amino acids.

著者

Yoshinori INOUE Atsushi YAMAMOTO

出版者

クロマトグラフィー科学会

雑誌

CHROMATOGRAPHY (ISSN:13428284)

巻号頁・発行日

vol.35, no.2, pp.63-72, 2014-08-10 (Released:2014-08-26)

参考文献数

94

被引用文献数

6

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High-performance liquid chromatography (HPLC) has great contribution in the analysis of the carbohydrates. Various separation modes such as ion exchange chromatography, size exclusion chromatography and partition chromatography have been applied to HPLC of mono- and oligosaccharides. Among them, partition chromatography with a polar stationary phase and aqueous mobile phase, called aqueous normal phase partition (ANP) chromatography, has been used in various fields, and is widely used for the separation of reducing sugars. Currently, this separation mode is considered to be a type of "hydrophilic interaction chromatography (HILIC)". As the use of HILIC mode increases, many novel stationary phases being available for the separation of reducing sugars have been developed. This article focuses on the separation of reducing sugars by ANP (HILIC) mode, and summarizes the traditional and/or recent stationary phases for the separation of reducing sugars. The applicability of various kinds of stationary phases to solid-phase extraction (SPE) of saccharides is also described.