Quantification of Type I Collagen α1 in Culture Medium Using Nano-Liquid Chromatography Tandem Mass Spectrometry

Ao GONG; Kazuo YAMADA; Li JING; Haruo TAKESHITA; Ken-ichi MATSUMOTO

doi:10.51010/sjms.38.1_1

Original Article

Quantification of Type I Collagen α1 in Culture Medium Using Nano-Liquid Chromatography Tandem Mass Spectrometry

Ao GONG, Kazuo YAMADA, Li JING, Haruo TAKESHITA, Ken-ichi MATSUMOTO

Author information

Ao GONG
Department of Biosignaling and Radioisotope Experiment, Interdisciplinary Center for Science Research, Organization for Research and Academic Information, Shimane University, Izumo, Shimane 693-8501, Japan
Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo, Shimane 693-8501, Japan
School of Basic Medical Science, Ningxia Medical University, Yinchuan, Ningxia 750004, China
Kazuo YAMADA
Department of Biosignaling and Radioisotope Experiment, Interdisciplinary Center for Science Research, Organization for Research and Academic Information, Shimane University, Izumo, Shimane 693-8501, Japan
Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo, Shimane 693-8501, Japan
Li JING
School of Basic Medical Science, Ningxia Medical University, Yinchuan, Ningxia 750004, China
Haruo TAKESHITA
Department of Legal Medicine, Faculty of Medicine, Shimane University, Izumo, Shimane 693-8501, Japan
Ken-ichi MATSUMOTO [Japan] Corresponding author
Department of Biosignaling and Radioisotope Experiment, Interdisciplinary Center for Science Research, Organization for Research and Academic Information, Shimane University, Izumo, Shimane 693-8501, Japan

Keywords: type I collagen, COL1A1, nano-LC/MS/MS, stable isotope-labeled peptide

JOURNAL FREE ACCESS FULL-TEXT HTML

2021 Volume 38 Issue 1 Pages 1-5

DOI https://doi.org/10.51010/sjms.38.1_1

Details

Abstract

Type I collagen, which consists of two α1 and one α2 chains, is an abundant extracellular matrix (ECM) protein. Changes in type I collagen are often associated with various diseases including fibrosis, osteogenesis imperfecta (OI), and Ehlers-Danlos syndrome (EDS). In the present study, we developed a method for quantification of type I collagen α1 (COL1A1) in a culture medium of LX-2 human hepatic stellate cells by using nano-liquid chromatography tandem mass spectrometry (nano-LC/MS/MS). After selecting a specific peptide of COL1A1, unlabeled and stable isotope-labeled peptides were used for quantitative analysis of COL1A1 by nano-LC/MS/MS. The concentration of secreted COL1A1 in the LX-2 cell culture medium was 38.78 ng/mL. The results indicate that this method is useful for quantifying COL1A1 in a cell culture medium.

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