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  1. Artikel ; Online: Proteomic Analysis of Adenovirus 5 by UHPLC-MS/MS: Development of a Robust and Reproducible Sample Preparation Workflow.

    Zarei, Mostafa / Jonveaux, Jérôme / Wang, Peng / Haller, Friedrich M / Gu, Bingnan / Koulov, Atanas V / Jahn, Michael

    ACS omega

    2022  Band 7, Heft 41, Seite(n) 36825–36835

    Abstract: Adenoviruses (AdVs) have recently become widely used therapeutic vectors for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine. AdVs are large, nonenveloped viruses with an icosahedral capsid formed from several proteins that encloses ... ...

    Abstract Adenoviruses (AdVs) have recently become widely used therapeutic vectors for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccine. AdVs are large, nonenveloped viruses with an icosahedral capsid formed from several proteins that encloses double-stranded DNA. These proteins are the main components and key players in initial stages of infection by the virus particles, so their heterogeneity and content must be evaluated to ensure product and process consistency. Peptide mapping can provide detailed information on these proteins, e.g., their amino acid sequences and post-translational modifications (PTMs), which is crucial for the development and optimization of the manufacturing processes. However, sample preparation remains the main bottleneck for successful proteomic analysis of the viral proteins (VPs) of AdVs due to their low concentrations and vast stoichiometric ranges. To address this problem, we have developed a fast and efficient protocol for preparing samples for proteomic analysis of VPs of AdV5 that requires no cleaning step prior to liquid chromatography-tandem mass spectrometry (LC-MS/MS). The approach enabled identification of 92% of amino acids in AdV5 VPs on average and quantification of 53 PTMs in a single LC-MS/MS experiment using trypsin protease. The data obtained demonstrate the method's potential utility for supporting the development of novel AdV-based gene therapy products (GTPs).
    Sprache Englisch
    Erscheinungsdatum 2022-10-05
    Erscheinungsland United States
    Dokumenttyp Journal Article
    ISSN 2470-1343
    ISSN (online) 2470-1343
    DOI 10.1021/acsomega.2c05325
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  2. Artikel ; Online: A novel protocol for in-depth analysis of recombinant adeno-associated virus capsid proteins using UHPLC-MS/MS.

    Zarei, Mostafa / Wang, Peng / Jonveaux, Jérôme / Haller, Friedrich M / Gu, Bingnan / Koulov, Atanas V / Jahn, Michael

    Rapid communications in mass spectrometry : RCM

    2022  Band 36, Heft 6, Seite(n) e9247

    Abstract: Rationale: In-depth characterization of the three capsid viral proteins (VPs 1, 2, and 3) of adeno-associated viruses (AAVs) is immediately needed to ensure the consistency in gene therapy products and processes. These proteins are typically present at ... ...

    Abstract Rationale: In-depth characterization of the three capsid viral proteins (VPs 1, 2, and 3) of adeno-associated viruses (AAVs) is immediately needed to ensure the consistency in gene therapy products and processes. These proteins are typically present at very low concentrations in matrices containing high concentrations of excipients and salts. Thus, there is a need for convenient methods for sample preparation before proteomic analysis. The aim of this study was to meet this need by developing a fast, reliable approach for isolating VPs in a manner enabling their efficient digestion and in-depth characterization using liquid chromatography-mass spectrometry (LC-MS).
    Methods: VPs from Anc80 were precipitated with different organic solvents, and the resulting precipitates were dissolved in either sodium deoxycholate (SDC) and N-dodecyl-beta-D-maltoside (DDM) or guanidine hydrochloride (Gu-HCl). The peptides obtained by the following enzymatic digestion by either trypsin or Asp-N were analyzed using LC-MS/MS.
    Results: We found that precipitation with chloroform/methanol/water results in fast, efficient preparation of VP samples, allowing 100% and 99.2% amino acid sequence coverage of VP1 for trypsin and Asp-N digestion, respectively. This also allowed complete sequence confirmation of VP1, VP2, and VP3 of Anc80, as well as characterization of the amino acid sequences of the N- and C-terminal regions of each VP, together with their post-translational modifications (PTMs).
    Conclusions: The presented method enables fast, reliable, and relatively cheap sample preparation for identifying AAV serotypes and characterizing the heterogeneity of capsid viral proteins, including their PTMs.
    Mesh-Begriff(e) Amino Acid Sequence ; Capsid/chemistry ; Capsid/metabolism ; Capsid Proteins/chemistry ; Capsid Proteins/genetics ; Capsid Proteins/metabolism ; Chromatography, High Pressure Liquid/methods ; Dependovirus/chemistry ; Dependovirus/genetics ; Dependovirus/metabolism ; Proteomics ; Recombinant Proteins/chemistry ; Recombinant Proteins/genetics ; Recombinant Proteins/metabolism ; Tandem Mass Spectrometry/methods
    Chemische Substanzen Capsid Proteins ; Recombinant Proteins
    Sprache Englisch
    Erscheinungsdatum 2022-01-10
    Erscheinungsland England
    Dokumenttyp Evaluation Study ; Journal Article
    ZDB-ID 58731-x
    ISSN 1097-0231 ; 0951-4198
    ISSN (online) 1097-0231
    ISSN 0951-4198
    DOI 10.1002/rcm.9247
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  3. Artikel: Optimisation of ITK inhibitors through successive iterative design cycles

    Herdemann, Matthias / Weber, Alexander / Jonveaux, Jérôme / Schwoebel, Frank / Stoeck, Michael / Heit, Isabelle

    Bioorganic & medicinal chemistry letters. 2011 Mar. 15, v. 21, no. 6

    2011  

    Abstract: Based on a hit cluster of compounds inhibiting interleukin-2 inducible T-cell kinase (ITK) in the submicromolar range a series of ITK inhibitor libraries were synthesized. Through iterative design cycles including kinase crystal structure information, ... ...

    Abstract Based on a hit cluster of compounds inhibiting interleukin-2 inducible T-cell kinase (ITK) in the submicromolar range a series of ITK inhibitor libraries were synthesized. Through iterative design cycles including kinase crystal structure information, indolylindazole libraries were identified which showed low nanomolar activity in enzymatic and cellular assays. The potential of these novel lead series was confirmed through in vivo tests in an anti-CD3-IL2 mouse model. The intravenous administration of highly potent ITK inhibitor 11o resulted in dose-dependent, efficient suppression of IL-2.
    Schlagwörter T-lymphocytes ; animal models ; crystal structure ; enzyme activity ; interleukin-2 ; intravenous injection
    Sprache Englisch
    Erscheinungsverlauf 2011-0315
    Umfang p. 1852-1856.
    Erscheinungsort Elsevier Ltd
    Dokumenttyp Artikel
    ZDB-ID 1063195-1
    ISSN 1464-3405 ; 0960-894X
    ISSN (online) 1464-3405
    ISSN 0960-894X
    DOI 10.1016/j.bmcl.2011.01.035
    Datenquelle NAL Katalog (AGRICOLA)

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  4. Artikel ; Online: Optimisation of ITK inhibitors through successive iterative design cycles.

    Herdemann, Matthias / Weber, Alexander / Jonveaux, Jérôme / Schwoebel, Frank / Stoeck, Michael / Heit, Isabelle

    Bioorganic & medicinal chemistry letters

    2011  Band 21, Heft 6, Seite(n) 1852–1856

    Abstract: Based on a hit cluster of compounds inhibiting interleukin-2 inducible T-cell kinase (ITK) in the submicromolar range a series of ITK inhibitor libraries were synthesized. Through iterative design cycles including kinase crystal structure information, ... ...

    Abstract Based on a hit cluster of compounds inhibiting interleukin-2 inducible T-cell kinase (ITK) in the submicromolar range a series of ITK inhibitor libraries were synthesized. Through iterative design cycles including kinase crystal structure information, indolylindazole libraries were identified which showed low nanomolar activity in enzymatic and cellular assays. The potential of these novel lead series was confirmed through in vivo tests in an anti-CD3-IL2 mouse model. The intravenous administration of highly potent ITK inhibitor 11o resulted in dose-dependent, efficient suppression of IL-2.
    Mesh-Begriff(e) Animals ; Crystallography, X-Ray ; Mice ; Models, Molecular ; Molecular Structure ; Protein Kinase Inhibitors/chemistry ; Protein Kinase Inhibitors/pharmacology ; Protein-Tyrosine Kinases/antagonists & inhibitors
    Chemische Substanzen Protein Kinase Inhibitors ; Protein-Tyrosine Kinases (EC 2.7.10.1) ; emt protein-tyrosine kinase (EC 2.7.10.2)
    Sprache Englisch
    Erscheinungsdatum 2011-03-15
    Erscheinungsland England
    Dokumenttyp Journal Article
    ZDB-ID 1063195-1
    ISSN 1464-3405 ; 0960-894X
    ISSN (online) 1464-3405
    ISSN 0960-894X
    DOI 10.1016/j.bmcl.2011.01.035
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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    Verfügbar in ZB MED Köln/Königswinter

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    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
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    ab Jg. 2022: Lesesaal (EG)

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  5. Artikel: Optimisation of ITK inhibitors through successive iterative design cycles

    Herdemann, Matthias / Weber, Alexander / Jonveaux, Jérôme / Schwoebel, Frank / Stoeck, Michael / Heit, Isabelle

    Bioorganic & medicinal chemistry letters

    Band v. 21,, Heft no. 6

    Abstract: Based on a hit cluster of compounds inhibiting interleukin-2 inducible T-cell kinase (ITK) in the submicromolar range a series of ITK inhibitor libraries were synthesized. Through iterative design cycles including kinase crystal structure information, ... ...

    Abstract Based on a hit cluster of compounds inhibiting interleukin-2 inducible T-cell kinase (ITK) in the submicromolar range a series of ITK inhibitor libraries were synthesized. Through iterative design cycles including kinase crystal structure information, indolylindazole libraries were identified which showed low nanomolar activity in enzymatic and cellular assays. The potential of these novel lead series was confirmed through in vivo tests in an anti-CD3-IL2 mouse model. The intravenous administration of highly potent ITK inhibitor 11o resulted in dose-dependent, efficient suppression of IL-2.
    Schlagwörter interleukin-2 ; intravenous injection ; enzyme activity ; T-lymphocytes ; animal models ; crystal structure
    Sprache Englisch
    Dokumenttyp Artikel
    ISSN 0960-894X
    Datenquelle AGRIS - International Information System for the Agricultural Sciences and Technology

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