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Host cell protein quantification workflow using optimized standards combined with data-independent acquisition mass spectrometry

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Journal of Pharmaceutical Analysis 2023年第5期13卷 494-502页

作者： Steve Hessmann Cyrille Chery anne-Sophie Sikora Annick Gervais christine Carapito BioOrganic Mass Spectrometry Laboratory(LSMBO) IPHC UMR 7178University of StrasbourgCNRSProFI-FR2048Strasbourg67000France Department of Analytical Development Sciences for Biologicals UCB Pharma S.A.Braine l'Alleud1420Belgium

Monitoring of host cell proteins(HCPs)during the manufacturing of monoclonal antibodies(mAb)has become a critical requirement to provide effective and safe drug ***-linked immunosorbent assays are still the gold standard methods for the quantification of protein ***,this technique has several limitations and does,among others,not enable the precise identification of *** this context,mass spectrometry(MS)became an alternative and orthogonal method that delivers qualitative and quantitative information on all identified ***,in order to be routinely implemented in biopharmaceutical companies,liquid chromatography-MS based methods still need to be standardized to provide highest sensitivity and robust and accurate ***,we present a promising MS-based analytical workflow coupling the use of an innovative quantification standard,the HCP Profiler solution,with a spectral library-based data-independent acquisition(DIA)method and strict data validation *** performances of the HCP Profiler solution were compared to more conventional standard protein spikes and the DIA approach was benchmarked against a classical datadependent acquisition on a series of samples produced at various stages of the manufacturing *** we also explored spectral library-free DIA interpretation,the spectral library-based approach still showed highest accuracy and reproducibility(coefficients of variation<10%)with a sensitivity down to the sub-ng/mg mAb ***,this workflow is today mature to be used as a robust and straightforward method to support mAb manufacturing process developments and drug products quality control.

关键词： Host cell proteins Absolute quantification standards Data-independent acquisition

Redox Regulation of Arabidopsis Mitochondrial Citrate Synthase

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Molecular Plant 2014年第1期7卷 156-169页

作者： Elisabeth Schmidtmann Ann-christine Konig anne Orwat Dario Leister Markus Hartl Iris Finkemeier Botany Department I of Biology Ludwig-Maximilians-University Munich Grosshaderner Strasse 2 82152 Planegg-Martinsried Germany

Citrate synthase has a key role in the tricarboxylic （TCA） cycle of mitochondria of all organisms, as it cata- lyzes the first committed step which is the fusion of a carbon-carbon bond between oxaloacetate and acetyl CoA. The regulation of TCA cycle function is especially important in plants, since mitochondrial activities have to be coordinated with photosynthesis. The posttranslational regulation of TCA cycle activity in plants is thus far almost entirely unexplored. Although several TCA cycle enzymes have been identified as thioredoxin targets in vitro, the existence of any thioredoxin-dependent regulation as known for the Calvin cycle, yet remains to be demonstrated. Here we have investigated the redox regulation of the Arabidopsis citrate synthase enzyme by site-directed mutagenesis of its six cysteine residues. Our results indicate that oxidation inhibits the enzyme activity by the formation of mixed disulfides, as the partially oxidized citrate synthase enzyme forms large redox-dependent aggregates. Furthermore, we were able to demonstrate that thioredoxin can cleave diverse intraas well as intermolecular disulfide bridges, which strongly enhances the activity of the enzyme. Activity measurements with the cysteine variants of the enzyme revealed important cysteine residues affecting total enzyme activity as well as the redox sensitivity of the enzyme.

关键词： citrate synthase mitochondria cysteine residues redox regulation thioredoxin TCA cycle Arabidopsis.

Pollinator diversity benefits natural and agricultural ecosystems,environmental health,and human welfare

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Plant Diversity 2022年第5期44卷 429-435页

作者： Daniel Mutavi Katumo Huan Liang anne christine Ochola Min Lv Qing-Feng Wang Chun-Feng Yang CAS Key Laboratory of Aquatic Botany and Watershed Ecology Wuhan Botanical GardenChinese Academy of SciencesWuhan 430074China College of Life Sciences University of Chinese Academy of SciencesBeijing 100049China Center of Conservation Biology Core Botanical GardensChinese Academy of SciencesWuhan 430074China

Biodiversity loss during the Anthropocene is a serious ecological *** are important vectors that provide multiple essential ecosystem services but are declining rapidly in this changing ***,several studies have argued that a high abundance of managed bee pollinators,such as honeybees(Apis mellifera),may be sufficient to provide pollination services for crop productivity,and sociological studies indicate that the majority of farmers worldwide do not recognize the contribution of wild pollinator diversity to agricultural ***,we review the importance of pollinator diversity in natural and agricultural ecosystems that may be thwarted by the increase in abundance of managed pollinators such as *** also emphasize the additional roles diverse pollinator communities play in environmental safety,culture,and *** indicates that in natural ecosystems,pollinator diversity enhances pollination during environmental and climatic perturbations,thus alleviating pollen *** agricultural ecosystems,pollinator diversity increases the quality and quantity of crop ***,studies indicate that many pollinator groups are useful in monitoring environmental pollution,aid in pest and disease control,and provide cultural and aesthetic *** the uncertainties that may accompany rapid environmental changes in the Anthropocene,the conservation of pollinator diversity must expand beyond bee ***,the value of pollinator diversity maintenance extends beyond the provision of pollination ***,conservation of pollinator diversity requires an interdisciplinary approach with contributions from environmentalists,taxonomists,and social scientists,including artists,who can shape opinions and behavior.

关键词： Bee Complementarity Ecological indicator Plant community Pollen limitation

Amplified fragment length polymorphism analysis supports the valid separate species status of Lucilia caesar and *** (Diptera: Calliphoridae)

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Forensic Sciences Research 2018年第1期3卷 60-64页

作者： christine J.Picard Jeffrey D.Wells anne Ullyot Knut Rognes Department of Biology&Forensic and Investigative Sciences Program Indiana University-Purdue University IndianapolisIndianapolisINUSA Department of Biological Sciences&International Forensic Research Institute Florida International UniversityMiamiFLUSA Biological Sciences University of ChicagoChicagoILUSA Department of Early Childhood Education Faculty of Arts and EducationUniversity of StavangerStavangerNorway

Common DNA-based species determination methods fail to distinguish some blow flies in the forensically and medically important genus Lucilia *** is a practical problem,and it has also been interpreted as casting doubt on the validity of some morphologically defined *** example is Lucilia illustris and ***,which co-occur in Europe whilst only *** has been collected in North *** that these species shared both mitochondrial and nuclear gene sequences,along with claims that diagnostic morphological characters are difficult to interpret,were used to question their separate species *** report here that amplified fragment length polymorphism profiles strongly support the validity of both species based on both assignment and phylogenetic analysis,and that traditional identification criteria based on male and female genital morphology are more reliable than has been claimed.

关键词： Forensic science forensic entomology Calliphoridae Lucilia caesar Lucilia illustris taxonomy AFLP analysis

Production and stability study of a hospital parenteral nutrition solution for neonates

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Journal of Pharmaceutical Analysis 2019年第2期9卷 83-90页

作者： anne-Laure Yailian Céline Serre Justine Fayard Marina Faucon Patrick Thomaré Samira Filali christine Pivot Florence Vételé Fabrice Pirot Emmanuelle Olivier Service Pharmaceutique Plateforme FripharmGroupe Hospitalier Centre Edouard HerriotHospices Civils de Lyon Laboratoire de Recherche et Développement de Pharmacie Galénique Industrielle UMR 5305Plateforme FripharmFaculté de PharmacieUniversité Claude Bernard Lyon 18 Service Pharmaceutique Site H?tel-DieuCentre Hospitalo-Universitaire de Nantes1place Alexis Ricordeau Service pharmaceutique Groupement Hospitalier EstHospices Civils de Lyon

Standard parenteral nutrition solutions are mixtures comprising interacting components that may degrade themselves over time. The objective of this study was to investigate the physicochemical and microbiological stability of a hospital preparation for parenteral nutrition in neonatology. The analyses were performed throughout the storage of the preparations at 2–8 °C(up to 4 months). The extent of stability was based on the determination of amino acids dosage, visual and physicochemical properties(glucose and electrolytes concentrations, pH and osmolality measurements, particle counting) and microbiological analysis(sterility test). A thermal degradation of ascorbic acid was conducted to evaluate the antioxidant properties of the parenteral mixture. Physicochemical and microbiological controls were found to comply with the specifications. Amino acids showed a good stability throughout the 4 months storage except for cysteine, which was progressively degraded to cystine, conferring a yellow coloration to parenteral solutions. Parenteral nutrition standards solutions remain stable for 4 months at 2–8 °C,ensuring safe administration in preterm infants.

关键词： Stability-indicating method study Parenteral nutrition Neonatology Amino acids High-performance liquid chromatographic

Long-term stability of gentamicin sulfate-ethylenediaminetetraacetic acid disodium salt (EDTA-Na_2) solution for catheter locks

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Journal of Pharmaceutical Analysis 2018年第6期8卷 386-393页

作者： anne-Sophie Fiolet Elise Jandot Pauline Doucey Coralie Crétet Célia Brunel christine Pivot Jean-Marc Ghigo Christophe Beloin David Lebeaux Fabrice Pirot Service Pharmaceutique Plateforme FRIPHARM Groupe Hospitalier Centre Edouard Herriot Hospices Civils de Lyon Unité de Génétique des Biofilms Département de Microbiologie Institut Pasteur Service de Microbiologie Unité Mobile de Microbiologie Clinique Assistance Publique-H?pitaux de Paris H?pital Européen Georges Pompidou Université Paris Descartes 12 rue de l'Ecole de Médecine Laboratoire de Recherche et Développement de Pharmacie Galénique Industrielle UMR 5305 Plateforme FRIPHARMF-69373 Faculté de Pharmacie Université Claude Bernard Lyon 1 8 avenue Rockefeller

A lock solution composed of gentamicin sulfate(5 mg/mL) and ethylenediaminetetraacetic acid disodium salt(EDTA-Na2, 30 mg/mL) could fully eradicate in vivo bacterial biofilms in totally implantable venous access ports(TIVAP). In this study, fabrication, conditioning and sterilization processes of antimicrobial lock solution(ALS) were detailed and completed by a stability study. Stability of ALS was conducted for12 months in vial(25 °C 7 2 °C, 60% 7 5% relative humidity(RH), and at 40 °C 7 2 °C, RH 75% 7 5%)and for 24 h and 72 h in TIVAP(40 °C 7 2 °C, RH 75% 7 5%). A stability indicating HPLC assay with UV detection for simultaneous quantification of gentamicin sulfate and EDTA-Na2 was developed. ALS was assayed by ion-pairing high performance liquid chromatography(HPLC) needing gentamicin derivatization, EDTA-Na2 metallocomplexation of samples and gradient mobile phase. HPLC methods to separate four gentamicin components and EDTA-Na2 were validated. Efficiency of sterility procedure and conditioning of ALS was confirmed by bacterial endotoxins and sterility tests. Physicochemical stability of ALS was determined by visual inspection, osmolality, pH, and sub-visible particle counting. Results confirmed that the stability of ALS in vials was maintained for 12 months and 24 h and 72 h in TIVAP.

关键词： Gentamicin-EDTA-Na2 loaded antimicrobial lock solution Pharmaceutical compounding Stability indicating HPLC assay method Totally implantable venous access ports

东非肯尼亚山植物多样性垂直格局和Rapoport海拔法则的检验

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东非肯尼亚山植物多样性垂直格局和Rapoport海拔法则的检验

作者： Ochola anne christine 中国科学院大学(中国科学院武汉植物园)

学位级别：硕士

物种丰富度沿海拔梯度的垂直格局研究是生物多样性及其保护等领域的一个重要方面。肯尼亚山是非洲第二高峰,为东非最为著名的一个死火山之一,但其在物种丰富度海拔格局和沿着海拔梯度的Rapoport海拔规则等方面的研究还比较缺乏。本研究... 详细信息

物种丰富度沿海拔梯度的垂直格局研究是生物多样性及其保护等领域的一个重要方面。肯尼亚山是非洲第二高峰,为东非最为著名的一个死火山之一,但其在物种丰富度海拔格局和沿着海拔梯度的Rapoport海拔规则等方面的研究还比较缺乏。本研究试图解释肯尼亚山植物的物种丰富度海拔格局,以及通过史蒂文斯方法(Stevens’ method)检测不同的植物组分(包括习性和地理分布)的Rapoport海拔法则。我们首先收集有关肯尼亚山植物分布的各种书籍和文献资料,以及鉴定肯尼亚国家博物馆植物标本馆的标本数据,同时结合我们多年的在肯尼亚山的考察结果,获得肯尼亚山的维管植物名录。我们将肯尼亚山海拔1200米至5000米,分为38个100米的海拔梯度,统计每个海拔带的物种多样性;同时我们将所有植物按植物的习性和地理分布分为12个不同组分,并使用史蒂文斯方法(Stevens’method)检测不同的植物组分的Rapoport海拔法则。我们的结果显示沿海拔梯度的植物物种丰富度呈现出驼峰状的分布,在中部海拔处丰富度达到峰值。年平均温度(AMT)是线性回归模型中最具影响力的解释变量,但物种丰富度也与面积密切相关。归一化的差异植被指数(NDVI)和年平均降水量(AMP)与物种丰富度的关系不大。尽管所有变量都可以解释驼峰模型,但AMT、面积和AMP最好地解释了物种丰富度的单峰模式以及中部海拔峰值。在海拔高度时,环境条件是合适的,特别是温度和降水,因此允许许多不同物种的生长和存活。相反,在高海拔地区,温度太低,导致物种丰富度下降,而在较低海拔时,由于环境温度,物种丰富度与较高海拔相比相对较高。尽管温度是影响最大的因素,但所有变量效应在丰富模式的形成中都是相互关联的。我们的研究结果量化了面积、AMT、AMP和NDVI对沿海拔梯度的植物物种丰富度分布格局的影响。在关于物种丰富度模式和海拔Rapoport规则(ERR)的研究中,我们的研究结果表明,与总物种的物种丰富度沿着海拔梯度显示出的驼峰形模式像是,不同的习性和地理分布的植物组分也显示出与总物种相似的驼峰形状。总物种和草本植物的平均海拔范围大小沿着海拔梯度呈现增加的模式,而在高海拔地区达到峰值后,木本植物和蕨类植物以及木本植物显示出明显的下降趋势。我们的结论是,广泛分布的草本植物种类也具有较宽的海拔范围,更适用于ERR,而在较高海拔地区出现的海拔范围较小的狭窄分布的木本植物可以逆转ERR。我们的研究结果表明,ERR在同一地区的不同生物组合并不一致。因此,该规则不是用于理解沿山地梯度的物种丰富度垂直模式的一致预测模式。

关键词：热带非洲肯尼亚山植物物种丰富度驼峰模型 Rapoport提升规则

Early Cretaceous Climate Changes Recorded in Magnetic Susceptibility and Color Index Variations of the Lower Liupanshan Group, Central China

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Acta Geologica Sinica(English Edition) 2016年第3期90卷 1011-1023页

作者： DAI Shuang ZHU Qiang HUANG Yongbo DA SILVA anne-christine ZHAO Jie LIU Junwei KONG Li PENG Dongxiang LUO Lingling YAN Ningyun WANG Wenjie ZHANG Xiang Key Laboratory of Western China's Environmental Systems(Ministry of Education) College of Earth and Environmental SciencesLanzhou University Cold and Arid Regions Environmental and Engineering Research Institute Chinese Academy of Sciences Shandong Institute of geological survey Pétrologie sédimentaire B20Allée du Six Ao t12Quartier AgoraUniversitéde Liège State Key Laboratory of Continental Dynamics Department of GeologyNorthwest University No.4 Institute of Geological & Mineral Resources Survey of Henan

This study carried out comprehensive analysis on sedimentology, magnetic susceptibility （7of） and color data of the continental sediments of the Liupanshan Group in Central China so as to obtain climatic change information during the 129.14-122.98 Ma interval. Based on the results of the Xlf and of the redness （a＊）, the section can be divided into two segments：（1） 129.14-126.3 Ma, with the lowest Xlf values and strongly variable relatively high values of redness and （2） 126.3-122.98 Ma, with high Elf values and relatively low redness. Analysis of the lithology and facies as well as the magnetic minerals and their contents points to a detrital origin of the magnetic minerals and this allow us to interpret the relationship between magnetic susceptibility variations and climate changes. Our study shows that the climate was significantly dry and hot during the whole studied interval although the interval between 126.3 Ma and 122.98 was a little bit cooler with increased humidity.

关键词： magnetic susceptibility color index climate change Liupanshan Group Early Cretaceous

The Outcome of Adolescent-Onset Anorexia Nervosa: A Study of 144 Cases

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Health 2014年第14期6卷 1883-1893页

作者： Solenn Kermarrec Bernard Kabuth anne-christine Rat Michel Vidailhet Colette Vidailhet EA 3444 Faculté de Médecine Université Henri Poincaré Nancy I Nancy France Pô le Hospital Universitaire de Psychiatrie de l’Enfant et l’adolescent Rennes France Service de Pédiatrie 3 et Génétique CHU Nancy Nancy France Service de Psychiatrie de l’Enfant et de l’Adolescent CHU Nancy Nancy France

Objectives: To evaluate the long-term outcome of adolescents with anorexia nervosa and attempt to define prognostic factors for outcome. Methods: Transversal, descriptive epidemiological study with a strict methodology: minimum interval of 4 years since first admission, evaluation using clearly defined diagnostic criteria (EAT-40 self-evaluation, HSCL, global patient-driven self-evaluation and hetero evaluation: clinical interview and LECE). The 198 subjects are former adolescent anorexics treated at the Hôpital d’Enfants (Children’s Hospital) in Nancy (France). Results: Clinical evaluation of the 144 subjects traced showed positive outcome for 68.1%, intermediate outcome for 23.2% and poor outcome for 8.7%. 5 factors for a good prognosis were found (middle socioeconomic class of parents, no prior treatment, hospitalization > 1 month,

关键词： Anorexia Nervosa Adolescent Epidemiology Prognosis Outcome