检索结果-内蒙古大学图书馆

Dissecting Methionine Oxidation by Hydrogen Peroxide in Proteins: Thermodynamics, Kinetics, and Susceptibility Descriptors

引用

JOURNAL OF CHEMICAL INFORMATION AND MODELING 2025年第2期65卷 749-761页

作者： De Sciscio, Maria Laura Centola, Fabio Saporiti, Simona D'Abramo, Marco Univ Rome Dept Chem I-00185 Rome Italy Merck Serono SpA Analyt Excellence & Program Management I-00012 Rome Italy

The oxidation of Met residues in proteins is a complex process, where protein-specific structural and dynamical features play a relevant role in determining the reaction kinetics. Aiming to a full-side perspective, we report here a comprehensive characterization of Met oxidation kinetics by hydrogen peroxide in a leptin protein case study. To do that, we estimated the reaction-free energy profile of the Met oxidation via a QM/MM approach, while the kinetics of the formation of the reactive species were calculated using classical molecular dynamics (MD) simulations. Our data, validated against the available experimental data on the Met oxidation in this protein, indicated that the protein's local and global motion represent the primary discriminating factor among residues' oxidation rates. Moreover, assuming that the free energy profile is independent of the specific protein system, the different reactivities of Met residues within five proteins (hGCSF, IL-1ra, leptin, somatotropin, and RNase) were qualitatively analyzed in terms of well-known structural/dynamic features, which can affect the kinetics of the whole process. The comprehensive analysis of the reaction thermodynamics and kinetics fingerprint enabled the identification of additional descriptors, helpful in assessing the susceptibility of protein-bound Met residues to oxidation.

关键词： Free energy

来源：评论

学校读者我要写书评

暂无评论

Exploring NIR spectroscopy data: A practical chemometric tutorial for analyzing freeze-dried pharmaceutical formulations

引用

CHEMOMETRICS AND INTELLIGENT LABORATORY SYSTEMS 2025年 257卷

作者： Massei, Ambra Cavallini, Nicola Savorani, Francesco Falco, Nunzia Fissore, Davide Politecn Torino Dipartimento Sci Applicata & Tecnol Corso Duca Abruzzi 25 I-10129 Turin Italy Merck Serono SpA Global Parenterals Dev Dept via Luigi Einaudi 11 I-00012 Rome Italy

Chemometrics tools are of fundamental importance for data analysis in the pharmaceutical field, especially with the increasingly strong assertion of the Process Analytical Technologies (PAT). In fact, analytical technologies such as Near-Infrared or Raman spectroscopies generate a lot of data, the spectra, that must be analyzed in a proper way. Typically, it is quite difficult to deeply understand the information hidden within the raw data. Therefore, careful, and efficient data exploration is needed to highlight the chemical and physical features of the analyzed samples. Here, a tutorial on all the fundamental steps and concepts needed to perform a proper data analysis based on a case-study of different freeze-dried formulations in the pharmaceutical field is proposed. The data analysis pipeline begins with the dataset explanation, to better point out the main known differences and similarities among the investigated formulations. After the first step of data preprocessing, Principal Component Analysis (PCA), Partial Least Squares (PLS) for regression, and Partial Least Squares-Discriminant Analysis (PLS-DA) for classification are presented and applied to show how to obtain deep comprehension of the real-case NIR dataset at hand. The experimental results demonstrate that trends related to increasing levels of sucrose and/or arginine, as well as distinct clusters related to the sample type and to the operator who conducted the analysis can be found and modelled in the example data. The tutorial aims at providing clear practical steps to conduct a robust data analysis, starting from the extraction and organization of the raw data, up to building more advanced predictive models (regression and classification). At each step some key questions are asked and answered to stimulate critical thinking in the reader. Also, commented MATLAB scripts are provided together with the real-case example NIR data, so that anyone could reproduce the whole data analysis in the tutorial,

关键词： Chemometrics Freeze-dry Near-infrared spectroscopy Exploratory analysis Regression analysis Classification analysis

来源：评论

学校读者我要写书评

暂无评论

NIR-Based Real-Time Monitoring of Freeze-Drying Processes: Application to Fault and Endpoint Detection

引用

PROCESSES 2025年第2期13卷 452-452页

作者： Massei, Ambra Falco, Nunzia Fissore, Davide Politecn Torino Dipartimento Sci Applicata & Tecnol Corso Duca Abruzzi 24 I-10129 Turin Italy Merck Serono SpA Global Parenteral Dev Via Luigi Einaudi 11 I-00012 Rome Italy

In the pharmaceutical industry, freeze-drying is crucial for the stability of active pharmaceutical ingredients (APIs). Monitoring this complex process presents challenges as traditional methods often lack real-time insights, potentially leading to quality issues and batch rejections. Effective monitoring is then essential for optimizing process parameters and minimizing waste, thus saving costs and resources. This study evaluated the application of Near-Infrared (NIR) spectroscopy for the real-time monitoring of the freeze-drying process: NIR spectra were acquired in-line via a specially designed flange in the freeze-dryer. Two approaches were investigated. The first involved freeze-drying monitoring using control charts, thus creating a reference model based on cycles under normal conditions. A PCA model was developed using these reference cycles, and an intentional fault cycle was performed to test the system's ability to detect deviations. Multivariate control charts, utilizing Hotelling's T2 and DModX metrics, were shown to effectively monitor process deviations, enhancing the understanding of freeze-drying dynamics. The second approach was based on the use of NIR spectroscopy for assessing residual moisture during lyophilization. By integrating Partial Least Squares (PLS) regression with in-line NIR spectra, we estimated endpoints and detected faults in both reference and faulty cycles. The results showed strong correlations between PLS estimates and the Pirani-Baratron method, highlighting NIR's applicability for monitoring drying phases. This research advocates for broader NIR implementation in pharmaceutical development, emphasizing its potential to monitor the process, ensure quality, and reduce out-of-specification product.

关键词： NIR spectroscopy process analytical technology freeze-drying biopharmaceuticals monitoring

来源：评论

学校读者我要写书评

暂无评论

Therapeutic management in women with a diminished ovarian reserve: a systematic review and meta-analysis of randomized controlled trials

引用

FERTILITY AND STERILITY 2025年第3期123卷 457-476页

作者： Conforti, Alessandro Carbone, Luigi Di Girolamo, Raffaella Iorio, Giuseppe Gabriele Guida, Maurizio Campitiello, Maria Rosaria Ubaldi, Filippo Maria Rienzi, Laura Vaiarelli, Alberto Cimadomo, Danilo Ronsini, Carlo Longobardi, Salvatore D'Hooghe, Thomas Esteves, Sandro C. Alviggi, Carlo Univ Naples Federico II Dept Neurosci Reprod Sci & Odontostomatol Via Sergio Pansini 5 I-80131 Naples Italy Univ Naples Federico II Dept Publ Hlth Naples Italy ASL Salerno Dept Obstet & Gynecol & Physiopathol Human Reprod Salerno Italy GENERA IVIRMA Global Res Alliance Clin Valle Giulia Rome Italy Univ Urbino Carlo Bo Dept Biomol Sci Urbino Italy Univ Campania Luigi Vanvitelli Dept Woman Child & Gen & Specialized Surg Naples Italy Merck Serono SpA Rome Italy Merck KGaA Darmstadt Germany Univ Leuven KU Leuven Biomed Sci Grp Dept Dev & Regenerat Leuven Belgium Androl & Human Reprod Clin ANDROFERT Campinas SP Brazil Aarhus Univ Fac Hlth Aarhus Denmark

Importance: The clinical management of women with diminished ovarian reserve (DOR) is a challenge in the field of medically assisted reproduction. Several therapeutic strategies have been proposed, but with mixed results, mainly because the definition of DOR used was inconsistent among trials. Objective: To investigate adjuvant treatments and protocols involving only women with DOR according to POSEIDON (Patient-Oriented Strategies Encompassing IndividualizeD Oocyte Number) criteria. Data Sources: We conducted a systematic search using the MEDLINE (PubMed), EMBASE, and ISI Web of Knowledge databases to identify relevant studies published up to June *** review protocol was registered at http://***/PROSPERO/ (registration number: CRD42022346117). Study Selection and Synthesis: After duplication removal, the titles and abstracts of 4,806 articles were scrutinized, and 124 full-text articles were assessed for eligibility. In total, 38 randomized controlled trials were included in the qualitative/quantitative analysis. The following interventions were evaluated: dehydroepiandrosterone (n- 1,336);testosterone (n- 418);high- vs. low-dose gonadotropin (n- 957);delayed-start protocol with gonadotropin hormone-releasing hormone antagonist (n- 398);letrozole (n- 612);clomiphene citrate (1,113);growth hormone (311);luteal phase stimulation (n- 57);dual triggering (n- 139);dual stimulation (168);luteinizing hormone (979);oestradiol pretreatment (n- 552);and corifollitropin alfa (n- 561). Main Outcomes: The primary outcome was live birth rate or ongoing pregnancy if data on live birth were unavailable. Secondary outcomes were number of oocytes retrieved, number of metaphase II oocytes, clinical pregnancy rate and miscarriage rate. Results: Testosterone supplementation is associated with higher live birth rates compared with nonsupplemented women among all interventions evaluated (odds ratio: 2.19, 95% confidence interval [CI]: 1.11-4.32, four studies, 368 pati

关键词： Ovarian reserve ART POSEIDON criteria IVF diminished ovarian reserve poor response poor responders poor ovarian reserve

来源：评论

学校读者我要写书评

暂无评论

Up-regulated microRNAs in blastocoel fluid of human implanted embryos could control circuits of pluripotency and be related to embryo competence

引用

JOURNAL OF ASSISTED REPRODUCTION AND GENETICS 2025年 1-15页

作者： Battaglia, Rosalia Caponnetto, Angela Ferrara, Carmen Fazzio, Anna Barbagallo, Cristina Stella, Michele Barbagallo, Davide Ragusa, Marco Vento, Maria Elena Borzi, Placido Scollo, Paolo Carli, Luca Feichtinger, Michael Kasapi, Evangelia Tsakos, Elias Palini, Simone Sierka, Wojciech Pecorino, Basilio Campitiello, Maria Rosaria Ronsini, Carlo Purrello, Michele Valerio, Domenico Longobardi, Salvatore D'Hooghe, Thomas Di Pietro, Cinzia Univ Catania Dept Biomed & Biotechnol Sci Sect Biol & Genet G Sichel I-95123 Catania Italy Univ Enna Kore Dept Med & Surg I-94100 Enna Italy Cannizzaro Hosp IVF Unit Catania Italy Wunschbaby Inst Feichtinger Vienna Austria EmbryoClin IVF Thessaloniki Greece Cervesi Hosp IVF Unit Cattolica Italy Gyncentrum Sp Zelazna 1 PL-40851 Katowice Poland ASL Salerno Dept Obstet & Gynecol & Physiopathol Human Reprod Salerno Italy Univ Campania Luigi Vanvitelli Dept Woman Child & Gen & Specialized Surg Naples Italy Inst Genet Res IRG I-80143 Naples Italy Merck Serono SpA Global Clin Dev Rome Italy Merck KGaA Darmstadt Germany KU Leuven Univ Leuven Dept Dev & Regenerat Biomed Sci Grp Leuven Belgium

Purpose The paper aims to investigate the biological role of microRNAs secreted by preimplantation embryo into the blastocoel fluid and to detect a distinctive molecular signature for identifying embryos with the highest implantation potential. Methods We carried on a multicenter retrospective study involving five European IVF centers. We collected 112 blastocoel fluid samples from embryos on day 5 post-fertilization, cultured individually, along with data on blastocyst grade and embryo transfer outcomes. Using a custom TLDA Array, we compared the expression levels of 89 miRNAs between 33 fluids from high-quality implanted embryos and 30 fluids from high-quality not-implanted embryos. Expression differences were assessed using SAM and t-test. Additionally, correlation and function enrichment analysis and network construction were conducted to identify the biological roles of deregulated microRNAs. Results We identified six up-regulated microRNAs in the blastocoel fluid from implanted embryos, significantly and positively correlated across all samples (r >= 0.7;P <= 0.05). They could take part in pluripotency circuits, regulating and being regulated by transcription factors associated with stemness, cell growth, and embryo development. The ROC curve analysis confirmed the potential of these miRNAs as implantation classifiers. Conclusion The six miRNAs up-regulated in blastocoel fluid from implanted embryos may represent a functional molecular signature for evaluating blastocyst quality and identifying the most competent embryos. Their evaluation associated with non-invasive preimplantation genetic testing, integrating epigenetic and genomic analyses, could enhance implantation grade and allow for identification of the euploid embryo not able to implant.

关键词： Blastocoel fluid Embryo development Embryo implantation Human blastocyst MicroRNAs

来源：评论

学校读者我要写书评

暂无评论

The Formidable Challenge of Controlling High Mannose-Type N-Glycans in Therapeutic mAbs

引用

TRENDS IN BIOTECHNOLOGY 2020年第10期38卷 1154-1168页

作者： Mastrangeli, Renato Audino, Maria Concetta Palinsky, Wolf Broly, Herve Bierau, Horst Merck Serono SpA Affiliate Merck KgaA Darmstadt G CMC Sci & Intelligence Biotech Dev Programme Rome Italy Merck Serono SpA Affiliate Merck KgaA Darmstadt G Biotech Dev Programme Aubonne Switzerland Merck Serono SpA Affiliate Merck KgaA Darmstadt G Biotech Proc Sci Corsier Sur Vevey Switzerland

The clinical efficacy and safety of therapeutic monoclonal antibodies (mAbs) are significantly affected by their Fc-glycosylation profile. High mannose-type N-glycans (HM) affect efficacy (in terms of antibody-dependent cell cytotoxicity), pharmacokinetics, and stability. While in endogenous IgGs the HM levels are very low, they are significantly higher in marketed therapeutic mAbs. In order to meet the demands for late-phase clinical trial and market supply, process intensification is required. Since glycosylation profiles are sensitive to process variations and changes, controlling HM levels in robust manufacturing processes presents a formidable challenge and requires a thorough understanding of the cellular processes as well as the biotechnical aspects that govern the production of HM glycans.

关键词： monoclonal antibody biopharmaceuticals N-glycosylation high mannose glycans glycoengineering C-type lectin receptors

来源：评论

学校读者我要写书评

暂无评论

Administration of follitropin alfa and lutropin alfa combined in a single injection: a feasibility assessment

引用

REPRODUCTIVE BIOLOGY AND ENDOCRINOLOGY 2009年第1期7卷 1-5页

作者： Agostinetto, Rita Merck Serono SpA I-00012 Rome Italy

Background: Gonadotrophins are routinely administered in assisted reproductive technology (ART) treatment protocols. Recombinant human follicle-stimulating hormone (r-hFSH;follitropin alfa) and recombinant human luteinizing hormone (r-hLH;lutropin alfa) can be administered individually or in a fixed combination. The ability to vary the FSH to LH dose ratio in a single injection without compromising the bioactivity of either gonadotrophin or generating losses of active principle is important for physicians and patients alike. Methods: This study investigated whether follitropin alfa (GONAL-f (R)), as lyophilized powder for reconstitution or solution from the GONAL-f (R) (filled-by-mass [FbM]) Prefilled Pen, could be used to reconstitute Pergoveris TM (follitropin alfa/lutropin alfa 150 IU/75 IU) lyophilized powder. In Ratio Groups 1 and 2, the r-hFSH: r-hLH ratio was 3:1;in Ratio Groups 3 and 4, the ratios of r-hFSH: r-hLH were 5:1 and 8:1, respectively. The protein content and bioactivity of each mixed solution were evaluated. The r-hFSH and r-hLH content was determined using reverse-phase high performance liquid chromatography. The biological activity of r-hFSH and r-hLH was assessed using the Steelman-Pohley and Van Hell in vivo bioassays in rats, respectively. Results: Follitropin alfa/lutropin alfa 150 IU/75 IU lyophilized powder could be successfully mixed with follitropin alfa 75 IU FbM solution that was either reconstituted from lyophilized powder or injected directly from the prefilled pen to create solutions with ratios of r-hFSH and r-hLH from 3:1 to 8:1. The measured content of r-hFSH and r-hLH corresponded favourably with the target protein content in Ratio Groups 1-4. The in vivo target and measured bioactivity of r-hFSH and r-hLH were also closely matched in all Ratio Groups. Conclusion: Follitropin alfa lyophilized powder or solution can be accurately mixed with follitropin alfa/lutropin alfa 150 IU/75 IU lyophilized powder to enable physicians to adm

关键词： Luteinizing Hormone Assisted Reproductive Technology Plastic Syringe Ratio Group Measured Protein Content

来源：评论

学校读者我要写书评

暂无评论

Analytical identification of additional impurities in urinary-derived gonadotrophins

引用

REPRODUCTIVE BIOMEDICINE ONLINE 2009年第3期19卷 300-313页

作者： Bassett, R. Lispi, M. Ceccarelli, D. Grimaldi, L. Mancinelli, M. Martelli, F. Van Dorsselaer, A. Merck KGaA Merck Serono SA Geneva Darmstadt Germany Merck KGaA Merck Serono SpA Darmstadt Germany Lab Spect Masse Bioorgan Strasbourg France

Advances in proteomic technology have enabled contaminant proteins to be identified from complex protein mixtures. The purity of two purified urinary gonadotrophin products, human menopausal gonadotrophin (u-HMG) and human FSH (u-hFSH), was compared with a preparation of recombinant human FSH (r-hFSH). After separation by sodium dodecyl sulphate-polyacrylamide, I electrophoresis (SDS-PAGE), western blot analysis showed that the recombinant preparation contained only FSH, whereas the urine-derived preparations exhibited several non-FSH or LH-related bands. These urinary components were further investigated by a proteomic approach using two-dimensional SDS-PAGE followed by mass spectrometric identification. The proteomic approach detected a total of 23 non-gonadotrophin-related proteins, at variable levels in different batches of the urine-derived preparations. Of these, 16 co-purified proteins have not been previously reported to be present in urine-derived gonadotrophins. These results indicate that the process used to purify urinary gonadotrophins may not remove all non-gonadotrophin proteins. By using a comprehensive proteomic approach, it has been shown that the recombinant FSH preparation has greater purity than either of the urine-derived preparations.

关键词： gonadotrophins impurities quality r-hFSH u-hFSH u-HMG

来源：评论

学校读者我要写书评

暂无评论

Degradation of an Fc-Fusion Recombinant Protein by Host Cell Proteases: Identification of a CHO Cathepsin D Protease

引用

BIOTECHNOLOGY AND BIOENGINEERING 2009年第6期104卷 1132-1141页

作者： Robert, Flavie Bierau, Horst Rossi, Mara Agugiaro, David Soranzo, Thomas Broly, Herve Mitchell-Logean, Christine Merck Serono SA Corsier Sur Vevey Merck Serono Biotech Ctr CH-1809 Fenil Sur Corsier Switzerland Merck Serono SpA Ardea Italy Univ Grenoble 1 UFR Med & Pharm Grenoble La Tronche France

A host-cell-related proteolytic activity was identified in a recombinant Fc-fusion protein production process. This report describes the strategy applied to characterize and isolate the enzyme responsible for this degradation by combining cell culture investigation and dedicated analytical tools. After isolation and sequencing of the clipped fragment generated in post-capture material, enzymatic activity was traced in different culture conditions, allowing identification of viable CHO cells as the source of protease. Inhibitors and pH screenings showed that the enzyme belongs to an aspartic protease family and is preferably active at acidic pH. The protease was isolated by purification on a pepstatin A column and characterized as a protein related to cathepsin D. An additional metallo-protease inhibited by EDTA was identified with an optimum activity at neutral pH. This study is an example of how quality and stability of therapeutic recombinant molecules are strongly influenced by cell culture parameters. Biotechnol. Bioeng. 2009;104: 1132-1141. (C) 2009 Wiley Periodicals, Inc.

关键词： CHO cell culture protease Fc-fusion fragmentation fed-batch cathepsin D

来源：评论

学校读者我要写书评

暂无评论

Industrial Case Studies Demonstrating Applicability of ICH M7 Control Options 3 and 4 for Nitrosamine Control

引用

ORGANIC PROCESS RESEARCH & DEVELOPMENT 2025年第4期29卷 1152-1167页

作者： Urquhart, Michael W. Burns, Michael J. Bernardoni, Frank Clark, Hugh F. Crochard, Jean-Philippe De Benedetti, Alessandro Dirat, Olivier Fennell, Jared W. Gall, Malcolm A. Y. Galli, Marzia Hildbrand, Stefan Kallemeyn, Jeffrey M. Kuhl, Nadine Mack, Daniel J. Moessner, Christian Pascoe, David D. Pozzoli, Alessandro Risch, Philippe Roberts, Alastair J. Teasdale, Andrew Thiel, Oliver R. Tomlin, Paula Whitehead, Andrew GlaxoSmithKline Stevenage SG1 2NY Herts England Lhasa Ltd Leeds LS1 5PS W Yorkshire England Merck & Co Inc Rahway NJ 07065 USA F Hoffmann La Roche Ltd Pharma Res CH-4070 Basel Switzerland Merck Serono SpA Guidonia Italy Pfizer Global Regulatory Sci Regulatory CMC Advisory Off Sandwich CT13 9NJ England Eli Lilly & Co Synthet Mol Design & Dev Indianapolis IN 46285 USA AstraZeneca Chem Dev Pharmaceut Technol & Dev Operat Macclesfield SK10 2NA England F Hoffmann La Roche Ltd CH-4070 Basel Switzerland AbbVie Inc Proc Chem N Chicago IL 60064 USA Vertex Pharmaceut Inc Proc Chem Boston MA 02210 USA Idorsia Pharmaceut Ltd CH-4123 Allschwil Switzerland Amgen Inc Drug Subst Technol Synthet Cambridge MA 02142 USA GlaxoSmithKline Cork T12 E1EC Ireland

The assessment and control of potential mutagenic impurities (PMIs) within pharmaceutical products are managed in accordance with the ICH M7 guideline "Assessment and control of DNA reactive (mutagenic) impurities in pharmaceuticals to limit potential carcinogenic risk". This guidance highlights four control options that can be used to give assurance of control of PMIs to below a level of toxicological concern for the intended use. These control options range from testing to confirm that impurity levels within the active pharmaceutical ingredient or product are below an acceptable limit (ICH M7 Option 1 Control) to a control strategy that relies on process controls and scientific principles (e.g., purging) to demonstrate impurity presence to below a level of concern in lieu of analytical testing (option 4). While ICH M7 control option 4 is an established approach to justify that levels of a potential mutagenic impurity are below an acceptable limit, there have been health authority challenges that the use of ICH M7 control option 4 rationales is not appropriate for N-nitrosamines without included confirmatory analytical testing data to confirm absence. The reasons behind this lack of acceptance for ICH M7 control option 4 alone may include (i) a higher perceived potency for nitrosamines over other mutagenic impurities as they alert as part of the cohort of concern and (ii) inappropriate application of purge rationales such that, in some instances, confirmatory testing data highlighted higher levels for the impurity than had been predicted to be present. Through the inclusion of industry relevant case studies, this publication outlines that, while some nitrosamines may require control to lower levels than the ICH M7 threshold of toxicological concern, the concept of the ICH M7 option 4 control is scientifically justified when the properties for the nitrosamine are considered and an appropriate conservative purge rationale is applied.

关键词： ICH M7 Option 4 mutagenic impurities genotoxic impurities impuritycontrol nitrosamines case studies

来源：评论

学校读者我要写书评

暂无评论

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

请选择保存的检索档案：

请选择收藏分类：

通借通还

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：