• Title/Summary/Keyword: PTMS

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

  • Oh, Myung Jin;Seo, Nari;Seo, JungA;Kim, Ga Hyeon;An, Hyun Joo
    • Mass Spectrometry Letters
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    • v.12 no.3
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    • pp.85-92
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    • 2021
  • 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.

Experimental development of the epigenomic library construction method to elucidate the epigenetic diversity and causal relationship between epigenome and transcriptome at a single-cell level

  • Park, Kyunghyuk;Jeon, Min Chul;Kim, Bokyung;Cha, Bukyoung;Kim, Jong-Il
    • Genomics & Informatics
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    • v.20 no.1
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    • pp.2.1-2.11
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    • 2022
  • The method of single-cell RNA sequencing has been rapidly developed, and numerous experiments have been conducted over the past decade. Their results allow us to recognize various subpopulations and rare cell states in tissues, tumors, and immune systems that are previously unidentified, and guide us to understand fundamental biological processes that determine cell identity based on single-cell gene expression profiles. However, it is still challenging to understand the principle of comprehensive gene regulation that determines the cell fate only with transcriptome, a consequential output of the gene expression program. To elucidate the mechanisms related to the origin and maintenance of comprehensive single-cell transcriptome, we require a corresponding single-cell epigenome, which is a differentiated information of each cell with an identical genome. This review deals with the current development of single-cell epigenomic library construction methods, including multi-omics tools with crucial factors and additional requirements in the future focusing on DNA methylation, chromatin accessibility, and histone post-translational modifications. The study of cellular differentiation and the disease occurrence at a single-cell level has taken the first step with single-cell transcriptome and is now taking the next step with single-cell epigenome.

Phenyl modified silica sol-gel films for photonics (Photonic 재로로서 페닐실리카 코팅막의 특성)

  • Ahn, Bok-Yeop;Seok, Sang-Il;Kim, Joo-Hyeun;Lim, Mi-Ae
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2003.11a
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    • pp.131-131
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    • 2003
  • The advent of photonic technologies in the field of communications and data transmission has been heavily increasing the demand in integrated optical (IO) circuits capable of accomplishing not only simple tasks like signal, but also more sophisticated functions like all-optical signal routing or active multiplexing/demultiplexing. In the last decade, sol-gel technology has been widely used to prepare optical materials. Sol-gel processes show many promises for the development of low-loss, high-performance glass integrated optical circuits. However, crack formation is likely to occur during heat treatment in thick gel films. In order to overcome the critical thickness limitation, the organic-modified silicate has been widely used. In this case coating matrices have been prepared from the organo-silanes of T structures, acidic catalyst and the as-prepared gel films have been heat-treated below 200$^{\circ}C$ to avoid the crack formation and the degradation of organic components. However, the films prepared in the acidic condition and the low heat temperature make the films contain high OH groups which is the major optical loss function. In this work, C$\sub$6/H$\sub$5/SiO$\sub$1.5/ films were prepared on silicon substrate by sol-gel method using base catalyst in a PTMS/NH$_4$OH/H$_2$O/C$_2$H$\sub$5/OH system. The sol showed spinable viscosity at 50 wt% of solid content, and neglectable viscosity change with time. The films were crack-free and transparent after curing at 450 $^{\circ}C$, and highly condensed to minimize OH content in C$\sub$6/H$\sub$5/SiO$\sub$1.5/ networks. The effects of heat treatment of the films are characterized on the critical thickness, the chemical composition and the refractive indices by means of SEM, FT-IR, TGA, prism coupler, respectively.

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Characterization of Ag Nanowire Transparent Electrode Fabricated on PVDF Film (PVDF 필름 위에 제작된 고전도도 Ag 나노와이어 투명전극 특성 연구)

  • Ra, Yong-Ho;Park, Hyelim;An, Soyeon;Kim, Jin-Ho;Jeon, Dae-Woo;Kim, SunWoog;Lee, Mijai;Hwang, Jonghee;Lim, Tae Young;Lee, YoungJin
    • Journal of Sensor Science and Technology
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    • v.28 no.6
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    • pp.366-370
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    • 2019
  • In this study, we have successfully fabricated a highly conductive transparent electrode using Ag nanowires, based on piezoelectric polyvinylidene difluoride (PVDF) film, that can be applied as transparent and flexible speakers. The structural morphology of the Ag nanowires was confirmed by a detailed scanning electron microscopy. Ultraviolet-visible spectroscopy demonstrated that the transparent electrode fabricated by the Ag nanowires exhibited a transmittance of above 70%. The transparent electrode also showed very low sheet resistance with high flexibility. We have further developed an anti-oxidation coating layer by using a tetraethyl orthosilicate-poly trimethyloxyphenylsilane (TEOS-PTMS) slurry technique. It was confirmed that the transmittance and sheet resistance of the antioxidant film depends critically on the humidity of the film surface. We believe such Ag nanowire electrodes are a very promising next-generation transparent electrode technology that can be used in future flexible and transparent devices.

Effect of intermolecular interactions between CNTs and silane binders on the opto-electrical properties of SWNT/silane binder films (탄소나노튜브와 바인더의 상호작용이 탄소나노튜브/바인더 박막의 정전기적 특성에 미치는 영향)

  • Han, Joong-Tark;Kim, Sun-Young;Jeong, Hee-Jin;Jeong, Seung-Yol;Lee, Geon-Woong
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2009.04b
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    • pp.97-98
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    • 2009
  • Here, we describe a versatile strategy for precise control of the optoelectrical properties of the single walled carbon nanotube (SWNT)/silane binder hybrid films by noncovalent hybridization. Stable SWNT/silane binder solutions were prepared by direct mixing of high concentration CNT solutions and silane sol solutions. The critical binder content was determined by varying the amount of binder in the SWNT/binder solutions. A binder content of 50 wt% was used to prepare the other SWNT/binder solutions. This study demonstrates how the intermolecular interactions between the SWNTs and the silanes can affect the conductivity of the CNT/binder network films by characterizing the optoelectrical and Raman spectroscopic properties of the SWNT/silane films containing silane binders with various functional groups. The use of the PTMS binder with phenyl groups was found to be most effective in the fabrication of transparent and conductive films on glass substrates. Such a precise control of the optoelectrical properties of SWNT/binder films can be useful to fabricate the high performance conductive thin films, with ramifications for understanding the fundamental intermolecular interaction in carbon materials science.

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Quinetides: diverse posttranslational modified peptides of ribonuclease-like storage protein from Panax quinquefolius as markers for differentiating ginseng species

  • Zhao, Qiang;Bai, Yunpeng;Liu, Dan;Zhao, Nan;Gao, Huiyuan;Zhang, Xiaozhe
    • Journal of Ginseng Research
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    • v.44 no.5
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    • pp.680-689
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    • 2020
  • Background: Peptides have diverse and important physiological roles in plants and are ideal markers for species identification. It is unclear whether there are specific peptides in Panax quinquefolius L. (PQ). The aims of this study were to identify Quinetides, a series of diverse posttranslational modified native peptides of the ribonuclease-like storage protein (ginseng major protein), from PQ to explore novel peptide markers and develop a new method to distinguish PQ from Panax ginseng. Methods: We used different fragmentation modes in the LTQ Orbitrap analysis to identify the enriched Quinetide targets of PQ, and we discovered Quinetide markers of PQ and P. ginseng using ultrahigh-performance liquid chromatography-quadrupole time-of-flight mass spectrometry analysis. These "peptide markers" were validated by simultaneously monitoring Rf and F11 as standard ginsenosides. Results: We discovered 100 Quinetides of PQ with various post-translational modifications (PTMs), including a series of glycopeptides, all of which originated from the protein ginseng major protein. We effectively distinguished PQ from P. ginseng using new "peptide markers." Four unique peptides (Quinetides TP6 and TP7 as markers of PQ and Quinetides TP8 and TP9 as markers of P. ginseng) and their associated glycosylation products were discovered in PQ and P. ginseng. Conclusion: We provide specific information on PQ peptides and propose the clinical application of peptide markers to distinguish PQ from P. ginseng.

Applications and Developmental Prospect of Protein Microarray Technology (Protein Microarray의 응용 및 발전 전망)

  • Oh, Young-Hee;Han, Min-Kyu;Kim, Hak-Sung
    • KSBB Journal
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    • v.22 no.6
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    • pp.393-400
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    • 2007
  • Analysis of protein interactions/functions in a microarray format has been of great potential in drug discovery, diagnostics, and cell biology, because it is amenable to large-scale and high-throughput biological assays in a rapid and economical way. In recent years, the protein microarray have broaden their utility towards the global analysis of protein interactions on a proteome scale, the functional activity analysis based on protein interactions and post-translational modifications (PTMs), and the discovery of biomarkers through profiling of protein expression between sample and reference pool. As a promising tool for proteomics, the protein microarray technology has advanced outstandingly over the past decade in terms of surface chemistry, acquisition of relevant proteins on a proteomic level, and detection methods. In this article, we briefly describe various techniques for development of protein microarray, and introduce developmental state of protein microarray and its applications.