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한국질량분석학회> Mass Spectrometry Letters> Analysis of Low Molecular Weight Collagen by Gel Permeation Chromatography

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Analysis of Low Molecular Weight Collagen by Gel Permeation Chromatography

Hee-jin Yoo , Duck-hyun Kim , Su-jin Park , Kun Cho
  • : 한국질량분석학회
  • : Mass Spectrometry Letters 12권3호
  • : 연속간행물
  • : 2021년 09월
  • : 81-84(4pages)
Mass Spectrometry Letters

DOI


목차

Introduction
Experimental
Results
Discussion
Conclusions
Acknowledgments
References

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초록 보기

Collagen, which accounts for one-third of human protein, is reduced due to human aging, and much attention is focused on making collagen into food to prevent such aging. Gel permeation chromatography with Reflective Index (RI) detection (GPC/RI) was chosen as the most suitable instrument to confirm molecular weight distribution, and we explored the use of this technique for analysis of collagen peptide molecular sizes and distributions. Data reliability was verified by matrix-assisted laser desorption/ionization coupled to time-of-flight (MALDI-TOF) mass spectrometric analysis. The data were considered meaningful for comparative analysis of molecular weight distribution patterns.

UCI(KEPA)

I410-ECN-0102-2022-400-000721366

간행물정보

  • : 자연과학분야  > 화학
  • : KCI등재
  • : SCOPUS
  • : 계간
  • : 2233-4203
  • : 2093-8950
  • : 학술지
  • : 연속간행물
  • : 2010-2022
  • : 255


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1Aerosol Mass Spectrometer (AMS)-Based Real-Time Physicochemical Characterization of Atmospheric Aerosols

저자 : Donghwi Kim

발행기관 : 한국질량분석학회 간행물 : Mass Spectrometry Letters 13권 2호 발행 연도 : 2022 페이지 : pp. 27-34 (8 pages)

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Atmospheric aerosols have become a major environmental concern because of their adverse effects on human health, air quality, and climate change. Over the last few decades, several mass spectrometry (MS)-based techniques have been developed and applied in the field of atmospheric aerosol research. Particularly, real-time measurement of ambient aerosols using an aerosol mass spectrometer (AMS) has become one of the most powerful tools for aerosol chemistry. This review provides a brief description of AMS and its applications for understanding the physicochemical properties of atmospheric aerosols, as well as its sources and evolution processes.

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2Lipid N-formylation Occurs During Fixation with Formalin

저자 : Min Jung Kim , Heejin Lim , Muwoong Kim , Yuri Choi , Thy N. C. Nguyen , Seung Cheol Park , Kwang Pyo Kim , Junyang Jung , Min-sik Kim

발행기관 : 한국질량분석학회 간행물 : Mass Spectrometry Letters 13권 2호 발행 연도 : 2022 페이지 : pp. 35-40 (6 pages)

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Human tissues and organs can be preserved intact by fixation with formalin for the future analysis of biomolecules of interest. With the advances in high-throughput methods, numerous protocols have been developed and optimized to attain the most pathophysiological information out of biomolecules, including RNA and proteins, in formalin-fixed samples. However, there is no systematic study to examine the effects of formalin fixation on the lipidome of biological samples in a global fashion. In this study, we conducted a mass spectrometry-based analysis to survey the alteration in the lipidome of mice brains by fixation methods. A total of 308 lipids were quantitatively measured using triple quadrupole mass spectrometry. We found that most were unchanged after formalin fixation except for a few lipid classes such as phosphatidylethanolamine.

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1Chromosome-Centric Human Proteome Study of Chromosome 11 Team

저자 : Heeyoun Hwang , Jin Young Kim , Jong Shin Yoo

발행기관 : 한국질량분석학회 간행물 : Mass Spectrometry Letters 12권 3호 발행 연도 : 2021 페이지 : pp. 60-65 (6 pages)

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As a part of the Chromosome-centric Human Proteome Project (C-HPP), we have developed a few algorithms for accurate identification of missing proteins, alternative splicing variants, single amino acid variants, and characterization of function unannotated proteins. We have found missing proteins, novel and known ASVs, and SAAVs using LC-MS/MS data from human brain and olfactory epithelial tissue, where we validated their existence using synthetic peptides. According to the neXtProt database, the number of missing proteins in chromosome 11 shows a decreasing pattern. The development of genomic and transcriptomic sequencing techniques make the number of protein variants in chromosome 11 tremendously increase. We developed a web solution named as SAAvpedia for identification and function annotation of SAAVs, and the SAAV information is automatically transformed into the neXtProt web page using REST API service. For the 73 uPE1 in chromosome 11, we have studied the function annotaion of CCDC90B (NX_Q9GZT6), SMAP (NX_O00193), and C11orf52 (NX_Q96A22).

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2Characterization of the N-glycosylation of Recombinant IL-4 and IL-13 Proteins Using LC-MS/MS Analysis and the I-GPA Platform

저자 : Ju Yeon Lee , Jin-woong Choi , Sanghyeon Bae , Heeyoun Hwang , Jong Shin Yoo , Joo Eon Lee , Eunji Kim , Young Ho Jeon , Jin Young Kim

발행기관 : 한국질량분석학회 간행물 : Mass Spectrometry Letters 12권 3호 발행 연도 : 2021 페이지 : pp. 66-75 (10 pages)

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Interleukin-4 (IL-4) and IL-13 are cytokines secreted by immune cells. Cytokines induce the proliferation of macrophages or promote the differentiation of secretory cells. The initiation and progression of allergic inflammatory diseases, such as asthma, are dependent on cytokines acting through related receptor complexes. IL-4 and IL-13 are N-glycoproteins. Glycan structures in glycoproteins play important roles in protein folding, protein stability, enzymatic function, inflammation, and cancer development. Therefore, the glycan structure of IL-4 and IL-13 needs to be elucidated in detail for the development of effective therapies. We report the first attempt to characterize the site-specific N-glycosylation of recombinant IL-4 and IL-13 via liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. The tandem mass spectra of intact N-glycopeptides were identified using the Integrated GlycoProteome Analyzer (I-GPA) platform, which can automatically and rapidly analyze multiple N-glycopeptides, including their glycan composition and amino acid sequences. The recombinant IL-4 and IL-13 were identified with amino acid sequence coverages of 84% and 96%, respectively. For IL-4, 52 glycoforms on one N-glycosylation site were identified and quantified. In IL-13, 232 N-glycopeptides from three N-glycosylation sites were characterized, with the site Asn52 being the most extensively glycosylated (~80%). The complex glycans were the most abundant glycan on IL-4 and IL-13 (~96% and 91%, respectively), and the biantennary glycans were the most abundant in both recombinant IL-4 and IL-13 proteins.

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3Serum Proteomic Analysis of Scrub Typhus Patients for Screening Antigenic Proteins Originating from Orientia tsutsugamushi

저자 : Sang-yeop Lee , Sung Ho Yun , Geul Bang , Chang-seop Lee , Seung Il Kim

발행기관 : 한국질량분석학회 간행물 : Mass Spectrometry Letters 12권 3호 발행 연도 : 2021 페이지 : pp. 76-80 (5 pages)

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Scrub typhus is an acute febrile disease caused by the pathogenic bacterium Orientia tsutsugamushi, belonging to the Rickettsiaceae family. The shotgun proteomic analysis was performed using the sera of scrub typhus patients to identify the proteins having their origin in O. tsutsugamushi. Three different databases approaches were used for the identification of the proteomes. We identified the RsmD, an RNA methyltransferase as the commonly detected protein from all three approaches. This protein was not detected in the sera of healthy negative controls. We believe that this protein is a potential biomarker of Orientia tsutsugamushi present in the sera of scrub typhus patients.

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4Analysis of Low Molecular Weight Collagen by Gel Permeation Chromatography

저자 : Hee-jin Yoo , Duck-hyun Kim , Su-jin Park , Kun Cho

발행기관 : 한국질량분석학회 간행물 : Mass Spectrometry Letters 12권 3호 발행 연도 : 2021 페이지 : pp. 81-84 (4 pages)

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Collagen, which accounts for one-third of human protein, is reduced due to human aging, and much attention is focused on making collagen into food to prevent such aging. Gel permeation chromatography with Reflective Index (RI) detection (GPC/RI) was chosen as the most suitable instrument to confirm molecular weight distribution, and we explored the use of this technique for analysis of collagen peptide molecular sizes and distributions. Data reliability was verified by matrix-assisted laser desorption/ionization coupled to time-of-flight (MALDI-TOF) mass spectrometric analysis. The data were considered meaningful for comparative analysis of molecular weight distribution patterns.

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5Facile and Rapid Glycosylation Monitoring of Therapeutic Antibodies Through Intact Protein Analysis

저자 : Myung Jin Oh , Nari Seo , Junga Seo , Ga Hyeon Kim , Hyun Joo An

발행기관 : 한국질량분석학회 간행물 : Mass Spectrometry Letters 12권 3호 발행 연도 : 2021 페이지 : pp. 85-92 (8 pages)

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The therapeutic antibody drug market has experienced explosive growth as mAbs become the main therapeutic modality for a variety of diseases. Characterization of glycosylation that directly affects the efficacy and safety of therapeutic monoclonal antibodies (mAbs) is critical for therapeutics development, bioprocess system optimization, lot release, and comparability evaluation. The LC/MS approach has been widely used to structurally characterize mAbs, and recently attempts have been made to obtain comprehensive information on the primary structure and post-translational modifications (PTMs) of mAbs through intact protein analysis. In this study, we performed state-of-the-art LC/MS based intact protein analysis to readily identify and characterize glycoforms of various mAbs. Different glycoforms of mAbs produced in different expression cell lines including CHO, SP2/0 and HEK cells were monitored and compared. In addition, the comparability of protein molecular weight, glycoform pattern, and relative abundances of glycoforms between the commercialized trastuzumab biosimilar and the original product was determined in detail using the given platform. Intact mAb analysis allowed us to gain insight into the overall mAb structure, including the complexity and diversity of glycosylation. Furthermore, our analytical platform with high reproducibility is expected to be widely used for biopharmaceutical characterization required at all stages of drug development and manufacturing.

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6MALDI-MS: A Powerful but Underutilized Mass Spectrometric Technique for Exosome Research

저자 : Iqbal Jalaludin , David M. Lubman , Jeongkwon Kim

발행기관 : 한국질량분석학회 간행물 : Mass Spectrometry Letters 12권 3호 발행 연도 : 2021 페이지 : pp. 93-105 (13 pages)

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Exosomes have gained the attention of the scientific community because of their role in facilitating intercellular communication, which is critical in disease monitoring and drug delivery research. Exosome research has grown significantly in recent decades, with a focus on the development of various technologies for isolating and characterizing exosomes. Among these efforts is the use of matrix-assisted laser desorption ionization (MALDI) mass spectrometry (MS), which offers high-throughput direct analysis while also being cost and time effective. MALDI is used less frequently in exosome research than electrospray ionization due to the diverse population of extracellular vesicles and the impurity of isolated products, both of which necessitate chromatographic separation prior to MS analysis. However, MALDI-MS is a more appropriate instrument for the analytical approach to patient therapy, given it allows for fast and label-free analysis. There is a huge drive to explore MALDI-MS in exosome research because the technology holds great potential, most notably in biomarker discovery. With methods such as fingerprint analysis, OMICs profiling, and statistical analysis, the search for biomarkers could be much more efficient. In this review, we highlight the potential of MALDI-MS as a tool for investigating exosomes and some of the possible strategies that can be implemented based on prior research.

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7A Study on the Reduction of Inorganic Arsenic in Hijiki and Rice Using the Various Pretreatments and Inductively Coupled Plasma Mass Spectrometry

저자 : Sang-ho Nam , Dong-chan Lee

발행기관 : 한국질량분석학회 간행물 : Mass Spectrometry Letters 12권 3호 발행 연도 : 2021 페이지 : pp. 106-111 (6 pages)

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Several pretreatment methods have been developed to reduce the inorganic arsenic, which is known to be highly harmful to humans, among various arsenic species present in hijiki and rice. The pretreatment methods were selected and developed as methods that can be non-harmful even after treatment and easily applied. Hijiki was applied by two methods. One was soaking in water at room temperature for various durations and the other was boiling of it in water for a short period of time. Rice was soaked in water with different rice-to-water ratios for various durations. The most effective method that reduced the inorganic arsenic in hijiki was to repeat parboiling for 5 minutes twice, which led to 79% reduction of the inorganic arsenic in it. In the case of rice, soaking for 24 hours at the ratio of 1:5 (rice:water) resulted in 51% reduction of inorganic arsenic in rice.

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8Simultaneous Determination of α-Amanitin and β-Amanitin in Mouse Plasma Using Liquid Chromatography-High Resolution Mass Spectrometry

저자 : Young Yoon Bang , Min Seo Lee , Chang Ho Lim , Hye Suk Lee

발행기관 : 한국질량분석학회 간행물 : Mass Spectrometry Letters 12권 3호 발행 연도 : 2021 페이지 : pp. 112-117 (6 pages)

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α-Amanitin and β-amanitin are highly toxic bicyclic octapeptides responsible for the poisoning of poisonous mushrooms such as Amanita, Galerina, and Lepiota by inhibiting RNA polymerase II, DNA transcription, and protein synthesis. A sensitive, simple, and selective liquid chromatography-high resolution mass spectrometric method using parallel reaction monitoring mode was developed and validated for the simultaneous determination of α- and β-amanitin in mouse plasma to evaluate the toxicokinetics of α- and β-amanitin in mice. Protein precipitation of 5 μL mouse plasma sample with methanol as sample clean-up procedure and use of negative electrospray ionization resulted in better sensitivity and less matrix effect. The calibration curves for α- and β-amanitin in mouse plasma were linear over the range of 0.5-500 ng/mL. The intra- and inter-day coefficient of variations and accuracies for α- and β-amanitin at four quality control concentrations were 3.1-14.6% and 92.5-115.0%, respectively. The present method was successfully applied to the toxicokinetic study of α- and β-amanitin after an oral administration of α- and β-amanitin at 1.5 mg/kg dose to male ICR mice.

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9Study on Photodegradable Water-Soluble Compounds of Expanded Polystyrene

저자 : Seulgidaun Lee , Sunghwan Kim

발행기관 : 한국질량분석학회 간행물 : Mass Spectrometry Letters 12권 3호 발행 연도 : 2021 페이지 : pp. 118-124 (7 pages)

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Many previous studies have focused on revealing the harmfulness of microplastic particles, whereas very few studies have focused on the effects of chemicals, particularly photooxidation product. In this study, products of photodegradation from expanded polystyrene (EPS), compounds produced by photolysis by ultraviolet (UV) light, were investigated. EPS was directly irradiated and photolyzed using a UV lamp, and then the extracted sample was analyzed using high-resolution mass spectrometry (HRMS). Multiple ionization techniques, including electrospray ionization, atmospheric pressure chemical ionization, and atmospheric pressure photoionization, were used. In total, >300 compounds were observed, among which polystyrene monomer, dimer, and oxidized products were observed. In this work, the data presented clearly demonstrate that it is necessary to identify and monitor oxidized plastic compounds and assess their effect on the environment.

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