Dissertations / Theses on the topic 'Fast protein liquid chromatography'
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Jones, Karen Lorraine. "Analysis of ferredoxin and flavodoxin in Anabaena and Trichodesmium using fast protein liquid chromatography." PDXScholar, 1988. https://pdxscholar.library.pdx.edu/open_access_etds/3812.
Full textNguyen, Nhung Phuong. "Axial Ligand Mutant: H229A." Digital Archive @ GSU, 2008. http://digitalarchive.gsu.edu/honors_theses/1.
Full textKirk, John Daniel. "Particle beam LC/MS with fast atom bombardment." Thesis, Georgia Institute of Technology, 1990. http://hdl.handle.net/1853/27127.
Full textEdwardson, P. A. D. "High Performance Liquid Chromatography of polynucleotides and proteins." Thesis, London School of Hygiene and Tropical Medicine (University of London), 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.376534.
Full textSmid, Jerusa. "Poliformismos do gene da proteína príon celular em pacientes com doença de Alzheimer." Universidade de São Paulo, 2011. http://www.teses.usp.br/teses/disponiveis/5/5138/tde-24052011-142607/.
Full textINTRODUCTION: The polymorphism in the prion protein gene (PRNP) may influence non prion neurological diseases. Some reports associate Alzheimers disease (AD) and the polymorphic codon 129 of the PRNP. This association has not been studied in Brazilian population. In this study we aimed to describe the association between the polymorphisms of codon 129 of the PRNP and AD. METHODS: One hundred AD patients were evaluated in the Cognitive and Behavioral Neurology Unit and Cognitive Disorders Reference Center of the Hospital das Clínicas of the University of São Paulo School of Medicine, matched for 111 controls, regarding to the PRNP polymorphism and cognitive measures. The PRNP polymorphisms were analyzed using denaturing high-performance liquid chromatography (DHPLC). Analyzes stratifying by apoE genotype was performed. RESULTS: The distribution of the codon 129 polymorphisms were: 45.5% M/M, 42.4% M/V and 12.2% V/V in AD patients; 39.6% M/M, 50.5% M/V and 9.9% V/V in the control group (p=0.503). The 117 codon analysis revealed silent allelic variant in 5% of AD patients and 3% of controls (p=0.780). The octarepeat deletion occurred in 5% of AD and 4% of controls (p=0.738). All AD patients and controls were N171N. One AD patient had a point mutation at codon 180 (V180I). Logistic regression failed to confirm any association between AD cognitive performance and the codon 129 of PRNP, as well as in the control group. There was no association between the codon 129 genotypes and genotypes and AD according to the apoE stratification. CONCLUSIONS: There were no differences in the frequency of the codon 129 polymorphism between AD. control group, according to the codon 129 polymorphisms. A point mutation at the codon 180 (V180I) was diagnosed in one patient
Bian, Juan. "Liquid Chromatography and Mass Spectrometry Based Analytical Method Development Towards Fast and Sensitive Analysis." The Ohio State University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=osu1557242729134942.
Full textAnsong, Godfred. "Analysis of plant polyphenols by high performance liquid chromatography/mass spectrometry and protein binding." Oxford, Ohio : Miami University, 2004. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=miami1083081905.
Full textZhou, Feng. "Protein characterization by capillary isoelectric focusing electrophoresis, reversed phase liquid chromatography and mass spectrometry." Access to citation, abstract and download form provided by ProQuest Information and Learning Company; downloadable PDF file, 103 p, 2008. http://proquest.umi.com/pqdweb?did=1456289241&sid=4&Fmt=2&clientId=8331&RQT=309&VName=PQD.
Full textMuir, Matthew Stewart. "Proteomics of the ovine cataract." Diss., Lincoln University, 2008. http://hdl.handle.net/10182/792.
Full textAtkinson, Ian E. "Mass Spectrometric Analysis of Environmental Contaminants, Protein Structure and Expression." Cleveland State University / OhioLINK, 2008. http://rave.ohiolink.edu/etdc/view?acc_num=csu1231174291.
Full textZhao, Bei. "Comparison of Label and Label-free Quantitative Liquid Chromatography Tandem Mass Spectrometry for Protein Biomarker Discovery." The Ohio State University, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=osu1285089805.
Full textRazzaque, Musaab Aaqib. "Studies of porphyrin glycoconjugates and amino acid-, peptide- and protein- adducts by liquid chromatography-mass spectrometry." Thesis, University of Leicester, 2001. http://hdl.handle.net/2381/30756.
Full textHidayah, Siti Nurul [Verfasser]. "Improvement of liquid chromatography for analysis and purification of proteoforms via rational protein purification parameter screening and sample displacement chromatography / Siti Nurul Hidayah." Hamburg : Staats- und Universitätsbibliothek Hamburg Carl von Ossietzky, 2021. http://d-nb.info/1241743037/34.
Full textNguyen, Anh Mai. "New approaches to preparation of macroporous monoliths for use in liquid chromatography." Doctoral thesis, Umeå : Department of Chemistry, Umeå Univ, 2009. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-20890.
Full textHoller, Christopher J. "Purification of an acidic recombinant protein from transgenic tobacco." Thesis, Virginia Tech, 2007. http://hdl.handle.net/10919/32379.
Full textPolyelectrolyte precipitation with polyethyleneimine (PEI) was identified as an initial purification step for purifying acidic recombinant proteins from tobacco. Polyethyleneimine precipitation allowed for high recovery and concentration of the target protein while removing large amounts of impurities from the initial extract. At dosages of 700-800 mg PEI/g total protein, nearly 100% of the rGUS activity was precipitated with generally more than 90% recovered from the pellet. In addition, more than 60% of the native tobacco proteins were removed in the process, resulting in a purification factor near 4.
Recombinant GUS was further purified by a step of hydrophobic interaction chromatography (HIC) followed by a step of hydroxyapatite chromatography (HAC). The HIC step served to remove PEI and other contaminants such as nucleic acids that were accumulated during the precipitation step, while the HAC step served to separate rGUS from the remaining native tobacco proteins, most notably ribulose 1,5-bisphosphate carboxylase-oxygenase (Rubisco). Nearly 40% of the initial rGUS activity was recovered as a near homogeneous fraction based on SDS-PAGE analysis after the three step process.
The main steps used in this process are anticipated to be scalable and do not rely on affinity separations, making the process potentially applicable to a wide variety of acidic recombinant proteins expressed in tobacco as well as other leafy crops.
Master of Science
Cai, Yi. "Coupling Ambient Ionization Mass Spectrometry with Liquid Chromatography and Electrochemistry and Their Applications." Ohio University / OhioLINK, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1472816263.
Full textSun, Xiaobo. "Forensic Applications of Gas Chromatography/Mass Spectrometry, High Performance Liquid Chromatography--Mass Spectrometry and Desorption Electrospray Ionization Mass Spectrometry with Chemometric Analysis." Ohio University / OhioLINK, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1329517616.
Full textChang, Chih-Hsiang. "Proteomic studies on protein N-terminus and peptide ion mobility by nano-scale liquid chromatography/tandem mass spectrometry." Doctoral thesis, Kyoto University, 2021. http://hdl.handle.net/2433/263596.
Full textCHILAKALA, SUJATHA. "DEVELOPMENT OF LIQUID CHROMATOGRAPHY-MASS SPECTROMETRIC ASSAYS AND SAMPLE PREPARATION METHODS FOR THE BIOLOGICAL SAMPLE ANALYSIS." Cleveland State University / OhioLINK, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=csu1512927043412916.
Full textWINTERS, MICHAEL SHAWN. "PROBING PROTEIN-PROTEIN INTERACTIONS in vitro and in vivo WITH CYANOGEN." University of Cincinnati / OhioLINK, 2002. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1027090541.
Full textNukareddy, Praveena. "Quantification Of Mouse Cardiac Troponin I And Myosin Binding Protein C Phosphorylation By Liquid Chromatography-Mass Spectrometry (lc-Ms)." ScholarWorks @ UVM, 2018. https://scholarworks.uvm.edu/graddis/986.
Full textMiao, Zhixin. "Development and Applications of Liquid Sample Desorption Electrospray Ionization Mass Spectrometry (DESI-MS)." Ohio University / OhioLINK, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=ohiou1347559532.
Full textRamström, Margareta. "Analysis of Complex Biological Samples using Liquid Chromatography-Fourier Transform Ion Cyclotron Resonance Mass Spectrometry." Doctoral thesis, Uppsala University, Analytical Chemistry, 2005. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-5729.
Full textStudies of protein and peptide expression are vital in order to understand complex biological systems. As demonstrated in this thesis, on-line packed capillary liquid chromatography-Fourier transform ion cyclotron resonance mass spectrometry (LC-FTICR MS) is a useful analytical tool for such studies.
A proteomics method, based on global tryptic digestion and subsequent separation and detection of the peptides by LC-FTICR MS, was developed for qualitative analysis of body fluids. Initial experiments on cerebrospinal fluid (CSF) provided results that were comparable or superior to those achieved by more time- and sample-consuming techniques. The method was also successfully applied on plasma and amniotic fluid. One of the major challenges in proteomics is the broad dynamic range of proteins in biological matrices. The advantages of removing high-abundant components from CSF and plasma prior to MS were demonstrated.
In order to search for potential biomarkers, mass chromatograms of CSF from patients suffering from amyotrophic lateral sclerosis (ALS) and controls were compared using an in-house constructed pattern recognition program. ALS-specific patterns were observed, and four out of five unknown samples were correctly assigned. Alternative strategies to quantitatively compare two pools of samples rely on differential chemical labeling. The performance of one such method, quantification-using-enhanced-signal-tags, was investigated in complex sample analysis. The experimental intensity ratios were proven to be consistent with the prepared concentration ratios of abundant proteins in CSF.
Finally, the thesis reports on the first experiments where electron capture dissociation (ECD) was successfully incorporated in on-line LC-MS experiments. ECD and nozzle-skimmer fragmentation were applied to a sample of endocrine peptides extracted from mouse pancreatic islets. The two fragmentation methods provided complementary information. However, the method needs further optimization before it can be applied in the analysis of more complex samples, such as body fluids.
Wagner, Knut. "Development of a comprehensive on-line multidimensional high performance column liquid chromatography system for protein and peptide mapping with integrated size selective sample fractionation." [S.l.] : [s.n.], 2001. http://ArchiMeD.uni-mainz.de/pub/2001/0143/diss.pdf.
Full textPaiva, Marcelo Vitor de. "Otimiza??o e valida??o de m?todos anal?ticos para a determina??o simult?nea de tuberculost?ticos (4-FDC) por CLAE/DAD e CLUE/ DAD." Universidade Federal do Rio Grande do Norte, 2013. http://repositorio.ufrn.br:8080/jspui/handle/123456789/13477.
Full textTuberculosis is a serious disease, but curable in practically 100% of new cases, since complied the principles of modern chemotherapy. Isoniazid (ISN), Rifampicin (RIF), Pyrazinamide (PYR) and Chloride Ethambutol (ETA) are considered first line drugs in the treatment of tuberculosis, by combining the highest level of efficiency with acceptable degree of toxicity. Concerning USP 33 - NF28 (2010) the chromatography analysis to 3 of 4 drugs (ISN, PYR and RIF) last in average 15 minutes and 10 minutes more to obtain the 4th drug (ETA) using a column and mobile phase mixture different, becoming its industrial application unfavorable. Thus, many studies have being carried out to minimize this problem. An alternative would use the UFLC, which is based with the same principles of HPLC, however it uses stationary phases with particles smaller than 2 ?m. Therefore, this study goals to develop and validate new analytical methods to determine simultaneously the drugs by HPLC/DAD and UFLC/DAD. For this, a analytical screening was carried out, which verified that is necessary a gradient of mobile phase system A (acetate buffer:methanol 94:6 v/v) and B (acetate buffer:acetonitrile 55:45 v/v). Furthermore, to the development and optimization of the method in HPLC and UFLC, with achievement of the values of system suitability into the criteria limits required for both techniques, the validations have began. Standard solutions and tablets test solutions were prepared and injected into HPLC and UFLC, containing 0.008 mg/mL ISN, 0.043 mg/mL PYR, 0.030 mg.mL-1 ETA and 0.016 mg/mL RIF. The validation of analytical methods for HPLC and UFLC was carried out with the determination of specificity/selectivity, analytical curve, linearity, precision, limits of detection and quantification, accuracy and robustness. The methods were adequate for determination of 4 drugs separately without interfered with the others. Precise, due to the fact of the methods demonstrated since with the days variation, besides the repeatability, the values were into the level required by the regular agency. Linear (R> 0,99), once the methods were capable to demonstrate results directly proportional to the concentration of the analyte sample, within of specified range. Accurate, once the methods were capable to present values of variation coefficient and recovery percentage into the required limits (98 to 102%). The methods showed LOD and LOQ very low showing the high sensitivity of the methods for the four drugs. The robustness of the methods were evaluate, facing the temperature and flow changes, where they showed robustness just with the preview conditions established of temperature and flow, abrupt changes may influence with the results of methods
A tuberculose ? uma doen?a grave, por?m cur?vel em praticamente 100% dos casos novos, desde que obedecidos os princ?pios da moderna quimioterapia. S?o considerados f?rmacos de 1? linha no tratamento ? tuberculose: isoniazida, pirazinamida, etambutol e rifampicina. De acordo com USP 33 - NF28 (2010) as an?lises cromatogr?ficas para 3 dos 4 f?rmacos (isoniazida, pirazinamida e rifampicina) duram em m?dia 15 minutos e mais 10 minutos para a obten??o do 4? f?rmaco (etambutol) utilizando outra coluna, com outra mistura de fase m?vel, tornando esta aplica??o na pr?tica industrial desfavor?vel. Uma das alternativas ? utilizar o CLUE, o qual baseia-se nos mesmos princ?pios da CLAE, por?m utiliza fases estacion?rias com part?culas menores que 2 ?m. Dessa forma pretende-se com o presente estudo desenvolver e validar novos m?todos anal?ticos para determina??o simult?nea de tuberculost?ticos por CLAE/DAD e CLUE/DAD. Para isto, foi realizado um screening anal?tico, o qual verificou que ? necess?rio um gradiente de sistema de fase m?vel A (tamp?o acetato:metanol 94:6 v/v) e B (tamp?o acetato:acetonitrila 55:45 v/v). Posteriormente, ao desenvolvimento e otimiza??o do m?todo em CLAE e CLUE com a obten??o dos valores de adequabilidade do sistema dentro dos limites de aceita??es vigente para ambos as t?cnicas, as valida??es deram-se in?cio. Solu??es padr?es e solu??es testes dos comprimidos foram preparadas e injetadas no CLAE e CLUE, contendo isoniazida, pirazinamida, etambutol e rifampicina nas concentra??es de 0,008, 0,043, 0,030 e 0,016 mg.mL-1, respectivamente. A valida??o dos m?todos anal?ticos foram realizadas para: especificidade / seletividade, intervalos da curva anal?tica, linearidade, limite de detec??o, limite de quantifica??o, exatid?o, precis?o (repetibilidade, precis?o intermedi?ria) e robustez. Os m?todos foram adequados para determina??o dos 4 f?rmacos separadamente sem interfer?ncia nos demais. Precisos, devido ao fato de que os m?todos demonstraram que mesmo com varia??o de dias, al?m da repetibilidade, os valores ficaram dentro da faixa preconizada na legisla??o vigente. Lineares (R > 0,99), ou seja, os m?todos foram capazes de demonstrar que os resultados obtidos eram diretamente proporcionais ? concentra??o do analito na amostra, dentro de um intervalo especificado. Exatos, uma vez que os m?todos foram capazes de apresentar valores de coeficiente de varia??o e porcentagem de recupera??o dentro dos limites exigidos (98 a 102%). Os m?todos mostraram LD e LQ com n?veis baixos demonstrando que os m?todos possuem elevada sensibilidade aos quarto f?rmacos. A robustez foi avaliada frente ?s altera??es de temperatura e fluxo, onde os m?todos demonstraram-se robustos apenas nas condi??es previamente estabelecidas de temperatura e fluxo, altera??es bruscas podem acarretar influ?ncia nos resultados dos m?todos
Wieland, Christoph. "Charakterisierung und Modifizierung poröser Cellulosepartikel für die flüssige Hochleistungs-Chromatographie und ihr Einsatz zur Untersuchung von Protein-Wechselwirkungen." Doctoral thesis, Humboldt-Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2010. http://dx.doi.org/10.18452/16090.
Full textPorous bead cellulose is an interesting material for the application in aqueous size exclusion chromatography (SEC). Its good modifiability makes it furthermore to a perfect starting material for protein aggregation studies. A protein with huge practical importance is insulin. Misfolding and aggregation of insulin creates serious problems e.g. in drug delivery systems. Thereby it undergoes a change from alpha-helix to beta-sheet structure and forms insoluble fibrils. A back-folding with (toxic) fluorinated alcohols and fluorinated nanoparticles was already shown in literature. The approach for this work was that fluorine (CF3-) on a surface with high hydroxyl-content can induce the back folding of proteins like insulin. The purpose was to get stationary phases that can induce back folding and separation of proteins on a single column. At first a characterization of suitable cellulose beads with focus on different porosimetry methods was done. For the first time a combination of “classical” porosimetry methods (Hg-Intrusion; N2-Sorption) with SAXS and inverse SEC was applied for porous cellulose particles. A reversible shrinking of pores during drying process was shown. Immobilization of fluorine on the surface of cellulose beads was done by grafting of fluorinated acrylates via cer(IV)-redox-initiation and by polymer analogous reaction with fluorinated iodo alkanes. Homopolymer free graft-copolymerization was achieved, whereas no effect on pore structure was observed. The control of protein adsorption on surface by chemical stimuli was shown. Aggregation studies using SEC, DLS and SAXS showed that fluoro-modified cellulose beads do not delay insulin-aggregation due to strong adsorption effects. Though a significant aggregation delay for insulin with unmodified cellulose beads was discovered.
Wiberg, Henning. "Analytical Approaches to Neurodegenerative Disease Protein Aggregation." Licentiate thesis, KTH, Analytisk kemi, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-34027.
Full textTang, Jianhua. "Development of a Novel Gradient Chromatofocusing Tandem Mass Spectrometry Technique for the Determination of Cationic Compounds in Biofluids; Identification of Caspase 3 Cleavage Sites of NHE-1 by High Performance Liquid Chromatography-Mass Spectrometry." Cleveland State University / OhioLINK, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=csu1247344073.
Full textAbbaraju, Naga Vijayalaxmi. "Patterns of protein expression in tissues of the killifish, Fundulus heteroclitus and Fundulus grandis." ScholarWorks@UNO, 2011. http://scholarworks.uno.edu/td/113.
Full textAl, Matari Amira. "Development of new analytical methods for the analysis at the intact level of glycoforms of hCG and other gonadotropins by nano liquid chromatography hyphenated to high resolution mass spectrometry." Thesis, Université Paris sciences et lettres, 2021. http://www.theses.fr/2021UPSLS034.
Full textGlycosylation is the attachment of glycans to a protein, so-called a glycoprotein. It is one of the most common post-translational modifications and it impacts protein activity and stability. Human chorionic gonadotropin (hCG) is a highly glycosylated protein, composed of two subunits, hCGα and hCGβ, having in total four N- and four O-glycosylation sites, leading to a very high number of glycoforms. However, the glycosylation state of this hormone essential for pregnancy was associated with some pregnancy pathologies. For this reason, it is necessary to be able to analyze the hCG glycoforms present in biological samples. In this work, new analytical methods for the analysis at the intact level of hCG glycoforms by nano liquid chromatography (nanoLC) hyphenated with mass spectrometry (MS) were developed. The miniaturization of the separation allowed an increase in sensitivity and its coupling with a high mass accuracy and high mass resolution analyzer, an Orbitrap, allowed the detection and semi-quantification of several α-subunit glycoforms in hCG-based drugs, but also of another gonadotropin sharing the same α-subunit, the follicle-stimulating hormone (FSH). The optimization of both a preconcentration step in the nanoLC-MS set-up and of the mobile phase gradient and nature allowed the detection of hCGβ glycoforms for the first time and also of a higher number of hCGα glycoforms. Next this analytical method using either formic acid or trifluoroacetic acid as mobile phase additive was evaluated for the detection of the α- and β-subunits of different recombinant or urinary gonadotropin-based drugs. Finally, with the aim to determine hCG glycoforms in serum of pregnant women, i.e complex biological samples, rather than in concentrated and fairly pure gonadotropin-based drugs, it was necessary to develop a selective extraction step of hCG glycoforms before their analysis in nanoLC-MS. This is why, immunosorbents were synthesized by immobilizing two different anti-hCG antibodies on a solid sorbent. The primary experiments of hCG immunoextraction showed promising results
Kamble, Sharad R. "Molecular interactions in pharmaceutical preformulation and supramolecular complexes. Structural properties governing drug-plasma protein binding and investigation of amino acids co-crystals." Thesis, University of Bradford, 2018. http://hdl.handle.net/10454/16882.
Full textJeudy, Jérémy. "Quantification de biomarqueurs protéiques dans des matrices biologiques complexes par spectrométrie de masse : développements et applications." Thesis, Lyon 1, 2014. http://www.theses.fr/2014LYO10242/document.
Full textOver the past decade, interest in the use of biomarkers in clinical studies has greatly increased. Quantification of a candidate protein biomarker in complex samples (eg. plasma) requires targeted and multiplexed assays. Immunoassays are the gold standard for their quantification. However, with the need for targeted and multiplexed methods, recent developments in mass spectrometry (MS) make a viable alternative to ELISA. The present work has addressed the problems encountered in this type of proteomic studies, and solutions that can be explored to improve the workflow of candidate biomarker’s evaluation. Methionine peptides are generally avoided due to their susceptibility to oxidation. However, it seemed interesting to study how their endogenous modifications could affect biological processes. In a first time, apolipoproteins were dismissed as a potential biomarker of Alzheimer’s disease due to oxidation impact. In the same time, problems associated with biological sample collection and storage were highlighted. DBS (Dried Blood Spot) and Vivid device evaluation from a panel of 32 blood proteins has provided a first possible solution to overcome these troubles. Thereafter, a new peptide quantification method called MRM3 was used to overcome biological matrix complexity. Reliable level determinations of 2 plasma proteins (C-Reactive protein and TIMP-1) and 2 urinary proteins (aquaporin-2 and podocin) were obtained. To improve sensitivity and reduce analysis solvent costs, performances of a micro chromatography platform were compared to a narrow-bore platform. This study highlighted the significant impact of the matrix effect on the analytical process, requiring new strategie development. Finally, to reduce sample complexity, evaluation of wide pore solid-phase extraction cartridges has been achieved. A protocol was successfully developed to analyze enzymes contained in commercial laundry samples. Finally to optimize biological sample preparation time, heated-assisted digestion and online desalting step were successfully associated. Only few hours were then required for quantitative analysis
Figueiredo, Douglas Borges de. "Desenvolvimento do processo de purificação da proteína A de superfície de pneumococo do clado 4 (PspA4Pro)." Universidade de São Paulo, 2014. http://www.teses.usp.br/teses/disponiveis/87/87131/tde-22102018-152147/.
Full textPneumococcal surface protein A (PspA) is found in all Streptococcus pneumoniae strains and is a promising candidate to be used in new pneumococcal vaccines. A purification process for PspA4Pro which inicial steps were: cell disruption, precipitation of the homogenate with the cationic detergent cetyltrimethylammonium bromide (CTAB) and pellet removal by centrifugation. The chromatographic techniques tested were ion exchange (anionic and cationic), immobilized metal affinity, hydrophobic interaction and mix mode with hydrophobic and cationic ligands. Using CTAB precipitation, anion exchange chromatography, crioprecipitation in pH4.0 and cation exchange chromatography the PspA reached the required purity (>95%) with recovery between 14% and 33% . The process reached acceptable levels of endotoxin in the final product and the purified PspA4Pro was recognized by anti-PspA4 antibodies and manteined its activity and secondary structure.
Aguiar, Mike. "Applications of mass spectrometry in clinical chemistry and biomedical research." Thesis, McGill University, 2007. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=108330.
Full textClinical chemistry is a medical discipline whose aim is to diagnose and assess disease by analysis of biological specimens. Modem laboratories can perform several hundred different tests using many different methods developed over the last century. The classical, more traditional assays are typically labour-intensive, not multiplexed (only measure one analyte or disorder per assay), expensive, require a long turnaround time, and may not provide adequate sensitivity and specificity. Developments in mass spectrometry (MS) and related technologies over the last two decades have provided solutions for many if not all of these shortcomings. While MS based applications have not yet been widely implemented in clinical chemistry laboratories, current developments will encourage the replacement of traditional methods as well as the expansion of clinically diagnostic endpoints. Indeed, modem MS can be used to simultaneously analyze and quantitate multiple biomarkers in a single analysis. Currently, no other technique exists that can provide a comparable multiplexed analysis. In this thesis, current MS and related technologies were developed and applied to several important but distinct clinical chemistry applications. [...]
La chimie clinique est une discipline medicale qui a pour but de diagnostiquer la presence et la progression d'une maladie par l'analyse d'echantillons biologiques. Les laboratoires modemes peuvent executer des centaines d'analyses en utilisant plusieurs methodes developpees au courrant des cent demieres annees. Les essaisc1assiques, et plus traditionnels, sont souvent laborieux, non multiplexe (mesurent seulement un analyte par essai), cher, exige un long temps de rotation et risque de ne pas fournir une specificite adequate. Pendant les deux dernieres decennies, les developpements dans Ie domaine de la spectrometrie de masse (MS) et les technologies rattachees ont foumi des solutions a plusieurs, pour ne pas dire tous, manques retrouves dans les methodes d'analyse traditionnelles.
Gorityala, Shashank. "TARGETED AND UNTARGETED OMICS FOR DISEASE BIOMARKERS USING LC-MS." Cleveland State University / OhioLINK, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=csu1547093694357568.
Full textCollin, Olivier L. "Development of a Novel Tandem Mass Spectrometry Technique for Forensic and Biological Applications." View abstract, 2007. http://gateway.proquest.com/openurl?url_ver=Z39.88-2004&res_dat=xri:pqdiss&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&rft_dat=xri:pqdiss:3292877.
Full textSloupová, Klára. "Izolace čistých aminokyselin z pšeničných otrub." Master's thesis, Vysoké učení technické v Brně. Fakulta chemická, 2021. http://www.nusl.cz/ntk/nusl-449764.
Full textCarvalho, Rimenys Junior. "Produção e purificação de um fragmento recombinante da proteína A de superfície do clado 3 (PspA3) de Streptococcus pneumoniae em Escherichia coli." Universidade de São Paulo, 2009. http://www.teses.usp.br/teses/disponiveis/87/87131/tde-11022010-121523/.
Full textThe pneumococcal surface protein A (PspA) is indispensable for virulence of S. pneumoniae and it was the first choice as carrier for a new conjugated vaccine against S.pneumoniae. Hence, the purpose of this work was to develop an industrial production and purification process of a recombinant fragment PspA clade 3 (rfPspA3) in E. coli. Fed-batch cultivations in 5 L bioreactors with defined medium were carried out using glucose or glycerol as carbon sources. It was obtained 62 g/L of dry cell weight and 3 g/L of rfPspA3. Cells were disrupted with 96.7% of efficiency by high pressure continuous homogenizer. Centrifugation was defined for the clarification step. The sequence with Q- followed by IMAC-Sepharose yielded the best purity and recovery of rfPspA3 (81 and 70%, respectively). Cation exchange was chosen for the last chromatography. In conclusion, an industrial production and purification process was developed and rfPspA3 was obtained with high purity (90%).
Bin, Saeedan Abdulaziz S. A. "The role of MMP10 in non-small cell Lung cancer, and pharmacological evaluation of its potential as a target for therapeutic intervention. Investigation of the role of MMP10 in the tumour microenvironment of non-small cell lung cancer using gene, protein and mass spectrometry approaches to determine MMP10’s potential in drug development strategies." Thesis, University of Bradford, 2014. http://hdl.handle.net/10454/14070.
Full textFull text was made available at the end of the embargo period, 12th Dec 2019
Bin, Saeedan Abdulaziz Saad Abdulaziz. "The role of MMP10 in non-small cell lung cancer, and pharmacological evaluation of its potential as a target for therapeutic intervention : investigation of the role of MMP10 in the tumour microenvironment of non-small cell lung cancer using gene, protein and mass spectrometry approaches to determine MMP10's potential in drug development strategies." Thesis, University of Bradford, 2014. http://hdl.handle.net/10454/14070.
Full textSimon, Romain. "La quantification ciblée de protéines et peptides par chromatographie liquide couplée à la spectrométrie de masse en tandem : développements analytiques et applications." Phd thesis, Université Claude Bernard - Lyon I, 2012. http://tel.archives-ouvertes.fr/tel-00875965.
Full textSANTANA, PATRICIA M. "Expressão de tireotrofina humana em células de embrião de rim humano (HEK293)." reponame:Repositório Institucional do IPEN, 2016. http://repositorio.ipen.br:8080/xmlui/handle/123456789/26947.
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Neste trabalho foi transfectada uma linhagem de células embrionárias de rim humano (HEK293) com os genes das subunidades α e β da tireotrofina humana (hTSH), hormônio glicoproteico secretado pela hipófise. Após 5 dias de cultivo obteve-se uma concentração de hTSH no meio condicionado de 0,95μg/mL. O material foi concentrado e purificado utilizando uma estratégia envolvendo duas etapas, uma cromatografia de troca catiônica e uma cromatografia líquida de alta eficiência (HPLC) de fase reversa, que permitiu uma recuperação de 55% e uma pureza >90%. O produto purificado (hTSH-HEK) foi analisado e comparado a uma preparação comercial obtida em células CHO (hTSH-CHO) e a uma preparação hipofisária (hTSH-Pit). A identidade e a pureza do hTSH-HEK foram avaliadas por métodos físicoquímicos e imunológico (espectrometria de massa MALDI-TOF, HPLC de exclusão molecular e de fase reversa, SDS-PAGE e ensaio imunoradiométrico). A porção glicídica do hTSH-HEK foi avaliada pela análise do perfil dos N-glicanos e o comportamento biológico deste hormônio foi avaliado por bioensaio in vivo e estudo farmacocinético. As 3 preparações apresentaram pureza equivalente (97%) e a massa molecular relativa do hTSH-HEK foi 2,1% menor do que a do hTSH-CHO e 2,7% maior do que a do hTSH-Pit. A maior hidrofobicidade relativa, avaliada por RP-HPLC, foi a do hTSH-HEK. Os N-glicanos identificados no hTSH-HEK foram do tipo complexo, apresentando predominantemente estruturas tri-antenárias, enquanto no hTSH-CHO e no hTSH-Pit as estruturas bi-antenárias foram predominantes. Foram detectadas diferenças significativas relacionadas à composição dos carboidratos para estas preparações, um teor muito menor de ácido siálico e muito maior de fucose foram observados no hTSHHEK. Foi confirmada a atividade biológica das 3 preparações, sendo a bioatividade do hTSHHEK 39% e 16% inferior à do hTSH-CHO e hTSH-Pit, respectivamente. A meia-vida circulatória do hTSH-HEK foi menor (1,5 X) que a do hTSH-CHO e a do hTSH-Pit (1,2 X). De acordo com esses resultados o hTSH-HEK pode ser considerado uma alternativa viável para aplicações clínicas especialmente por sua origem humana e composição de carboidratos.
Dissertação (Mestrado em Tecnologia Nuclear)
IPEN/D
Instituto de Pesquisas Energéticas e Nucleares - IPEN-CNEN/SP
Arnell, Robert. "Development and Validation of Methods for Characterization of Multi-Component Systems in Preparative LC." Doctoral thesis, Uppsala : Acta Universitatis Upsaliensis, 2006. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-7422.
Full textPerchepied, Stan. "Nouveaux outils miniaturisés pour l’analyse de biomolécules dans des fluides biologiques." Thesis, Sorbonne université, 2020. https://accesdistant.sorbonne-universite.fr/login?url=http://theses-intra.upmc.fr/modules/resources/download/theses/2020SORUS105.pdf.
Full textProtein analysis is mainly carried out using the "bottom-up" approach which is based on enzymatic digestion of proteins and analysis of resulting peptides by liquid chromatography coupled with tandem mass spectrometry. Usually, enzymatic digestion is carried out in solution. However, this procedure is long. The use of immobilised enzyme reactors (IMER) and their hyphenation with LC-MS/MS increases the reliability and sensitivity of the overall method. The study conducted during this thesis had two distinct objectives. The first was to evaluate the potential of IMERs for glycosylation characterisation. The complementarity of IMERs of pepsin and trypsin, developed in classical format by grafting the proteases on a Sepharose support, allowed to identify the N-glycans on the 4 glycosylation sites a pregnancy hormone contained in 2 drugs. The second objective was the miniaturization of these tools. To do this, monoliths obtained by sol-gel approach in the presence of organosilanes or by radical polymerization of organic monomers were synthesized in situ in capillaries of 100 µm internal diameter. The better repeatability of synthesis of organic supports led to their selection for functionalization by trypsin or pepsin. In parallel, a miniaturised set-up for the analysis of digests was carried out. This will allow the subsequent inclusion of IMER in the overall analytical device
Palmblad, Magnus. "Identification and Characterization of Peptides and Proteins using Fourier Transform Ion Cyclotron Resonance Mass Spectrometry." Doctoral thesis, Uppsala universitet, Institutionen för materialvetenskap, 2002. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-1999.
Full textZanolini, Carolina. "Estabilidade de formulações cosméticas antienvelhecimento com hidrolisado de proteína do arroz (Oryza sativa)." Universidade de São Paulo, 2011. http://www.teses.usp.br/teses/disponiveis/9/9139/tde-22122011-083227/.
Full textThe peptides are being widely used in cosmetics, since they act on specific receptors in cells and organs. In particular the hydrolyzed rice (Oryza sativa) protein has the ability to increase the production of fibroblasts and collagen. This study aims to assess the stability of cosmetic formulations, with or without the presence of hydrolyzed samples of rice protein, by thermal and physical-chemical methods, the development of analytical methodologies for the separation and identification of samples of hydrolyzed rice protein and the determination of the antioxidant activity of these samples. Cosmetic formulations have been developed and studies of physical and chemical stability and heat resistance were carried out. The physical and organoleptic characteristics were also studied. It was observed that the approved formulations presented small pH variation. They were then studied with the incorporation of hydrolyzed rice protein samples, and approved. To determine the thermal stability of hydrolyzed rice protein samples in cosmetic preparations, analytical techniques as thermogravimetry / derivative thermogravimetry were used in this research. A method was developed to evaluate the antioxidant activity of hydrolyzed rice protein samples, which showed a variation of activity ± 4 times between samples. Methods have been developed by high performance liquid chromatography and high performance liquid chromatography coupled to mass spectrometry for separation and identification of peptides present in the studied samples containing hydrolyzed rice protein.
Chen, Rong-Chun, and 陳容峻. "Fast Determination of Phosphorylated Proteins in Biological Samples by Liquid Chromatography- Tandem Mass Spectrometry." Thesis, 2011. http://ndltd.ncl.edu.tw/handle/15225879427163346391.
Full text高雄醫學大學
生物化學研究所
99
Protein phosphorylation is one of post-translational modifications (PTMs) that served as a central mechanism in signal transduction of biological organisms. Approximately 99% of this modification located on serine and threonine residues, whereas a few occurred on tyrosine residue. However, due to the facility in designing epitopes of antibodies for phosphotyrosine, it facilitated the detection and functional analysis of phosphotyrosine in biological organisms by many methods (such as western blotting, immunoprecipitation, in vitro kinase assay and mass spectrometry). Mass spectrometry (MS) is one of the major methods for proteomics analysis. This study developed a convenient way for protein digestion by using microwave oven to shorten the digestion time. Protein sample volume was at microscale level (2μL). The results showed that microwave-assisted digestion had better efficiency than conventional method, and this strategy also shortened the digestion time from 16 hours to 2 minutes. After protein digestion, chemical derivatization was introduced for labeling phosphorylated proteins, which increased the ionization efficiency and decreased neutral loss of phosphate group in MS analysis. Moreover, relative quantification of phosphorylation on the same peptide could be calculated by comparing the homologue chemicals labeled on the peptide. In the third part of this study, the calibration curve of digested human serum albumin (HSA) was constructed by mass spectrometric multiple reaction monitoring (MRM) assay. HSA quantification was used as a benchmark for quantifying phosphorylated proteins in human blood and urine. In conclusion, a fast and convenient method for analyzing phosphorylated proteins by MS was established. The advantages of this method contain microscale samples, shorter digestion time, and good detection and quantification efficiency of phosphorylated proteins. Application of this method in biological samples demonstrated workable.
HSIAO, KAI YUN, and 蕭凱云. "Protein Separation on Reversed-Phase Capillary Liquid Chromatography." Thesis, 2000. http://ndltd.ncl.edu.tw/handle/47132463141535814083.
Full text國立成功大學
化學系
88
In this study, the parameters affecting protein separation on reversed-phase liquid chromatography, such as organic solvent、ion-pairing agent and the pressure were investigated. For the effect of pressure, the thermodynamic principals were applied to infer the cause of observed results. For organic solvent effect, three model proteins, ribonuclease A、lysozyme and α-lactalbumin which have the similar molecular weight and number of amino acids but different hydrophobicity, were judiciously chosen for this investigation. It was found that the separation efficiency using methanol as organic modifier is always worse than using acetonitrile. Moreover the efficiency difference between methanol and acetonitrile were smaller for the more hydrophobic proteins. It could be that the hydrophobic proteins were easily solvated by organic solvents such that less interaction with stationary phase once desorbed. For ion-pairing agent effect, two homologous series, poly-L-phenylalanine and poly-L-lysine, in addition to the three model proteins were used to investigate the effect of four ion-pairing agents : phosphoric acid、formic acid、trifluoroacetic acid and heptafluorobutyric acid. It was found that the retention time was increased with the hydrophobicity of ion-pairing agents. Moreover, the retention time of the hydrophilic peptides was more sensitive to the change of ion-pairing agents. The more hydrophilic peptides resulted in a better separation efficiency when paired with a more hydrophobic ion-pairing agent. Furthermore, the separation of lysozymes from chicken and turkey which are only different in 7 amino acids out of total 129 amino acids, had a better resolution and efficiency ehen under an isocratic elution using the mobile phase composed of acetonitrile and trifluoroacetic acid. For pressure effect, it was demonstrated that the retention time of the lysozyme was increased by as much as two to three times as the absolute pressure on the viewing window was increased from 23 to 318 bar. Since many factors such as conformational changes of proteins, ionization, and hydrophobic interactions may all contribute to the protein retention and are subjective to change with the pressure, it is desirable to find out the major cause of the pressure effect. In this study, a homologous series of poly-aminoacid which have no secondary structures was investigated. It was found that the more hydrophobic peptides resulted in a more pronounced pressure-induced retention and consequently, a greater volume change ( △V＝Vsta－Vmob ) which was calculated from the first derivative of the lnk'' versus pressure polts. For example, the volume change of hydrophobic poly-L-phanylalanine was determined to be around minus 10 cm3/mol per phenylalanine, and the volume change of hydrophilic poly-L-lysine is about minus 1.5 cm3/mol per lysine. It is believed that perturbations in solute ionization、conformation change and stationary phase structural change have a minor impact under the investigated conditions, and the pressure-induced shift of the equilibria regarding hydrophobic ad-desorption is the major cause of the observed increase of protein retention. Assuming a linear relationship between the volume change and the hydrophobicity of amino acids, the volume change of other amino acids could be estimated. Moreover from comparing the volume changes of lysozyme and phenylalanine, it was predicted that about tens phenylalanine-equivalent residues on the lysozyme surface were involved in the hydrophobic association with the chromatographic ligands.
(9179615), Yun Yang. "STUDY FOR THE MECHANISM OF PROTEIN SEPARATION IN REVERSED-PHASE LIQUID CHROMATOGRAPHY." Thesis, 2020.
Find full textLiquid chromatography coupling with mass spectrometry (LC/MS) plays an important role in pharmaceutical characterization because of its ability to separate, identify, and quantify individual compounds from the mixture. Polymer brush layer bonded to the silica surface is designed as a novel stationary phase to improve the LC resolution and MS compatibility. The polymer thickness can be controlled to shield the analyte from interacting with the active silanol on the surface and reduce peak tailing. The functional group of the polymer can be changed to tune the selectivity in different separation modes.
Two projects on LC/MS method development for biomolecule characterization using polymer-shell column are discussed in this work. In the first project, a polymer-shell column is used for disulfide bonds and free thiol subspecies identification, which is a major type of structural heterogeneities in IgG1. Compared with commercial columns, the polymer-shell column is able to resolve the free thiol variants without the presence of trifluoroacetic acid and greatly improve the MS signal. In the second project, a polymer-shell column is used for characterizing the drug-loading profile for antibody-drug-conjugates (ADC) via online LC/MS. The separation employs a mobile phase of 50 mM ammonium acetate to keep the ADC intact, and a gradient of water/isopropanol for ADC elution. MS data show that all ADC species remained intact and native on the column. Positional isomers can be separated and identified with the new method as well. Furthermore, to understand the surface chemistry and protein separation behavior quantitatively, a chromatographic simulation study is performed. The result shows that protein separation in RPLC can be described by a bi-Langmuir adsorption isotherm with mixed-mode retention of strong and weak sites. Smaller fractions and lower equilibrium constant of the strong site, which is the active silanol, give less tailing for protein separation.
Hou, Chun Tsen, and 侯春岑. "Effect of Pressure on Protein Retention in Reversed-Phase High-Performance Liquid Chromatography." Thesis, 1999. http://ndltd.ncl.edu.tw/handle/45187504867671626285.
Full text國立成功大學
化學系
87
Pressure-induced change of protein behavior has long been either ignored or observed to occur at high pressures (>1 kbar). In this study, the on-column detection system is used to directly investigated the effect of pressure on protein retention in situ along the column. The range of pressure in this study is from 20 bar to 360 bar. Meanwhile, in order to observe the tertiary structure change of lysozyme under pressure effect, the on-the-fly scan UV spectroscopy (A281nm/292nm) was acquired as well. In this work, three kinds of stationary phases, standard C18, C4 and low density C18 were investigated. The isocratic elution behavior of lysozyme molecule was investigated by the dual on column detection system before pressure study. From the relationship between capacity factor (k') and the compositions of mobile phase (acetonitrile or methanol), the desorbed and adsorbed conditions were defined. In the desorbed condition, the lysozyme molecule is hardly retained on the stationary phase (k'~0), whereas, in the adsorbed condition the solute molecule is strongly retained (k'>0.5). In this study, the chromatographic retention of lysozyme molecule was measured directly on a transparent fused silica column as a function of the local pressure under both desorbed and adsorbed conditions. Under the desorbed condition, both the capacity factor (k') and bandwidth of lysozyme are not changed significantly as the pressure increased. Under the adsorbed condition, the capacity factor (k') is substantially increased and the bandwidth is broadened upon pressurizing. From the thermodynamic theory, the molar volume change(ΔV=Vsta-Vmob) of the protein between stationary phase (Vsta) and mobile phase (Vmob) can be determined from the slope of the plot of lnk' versus pressure. The linear equation was attempted to fit the plots constructed under adsorbed conditions, resulting in R2 values greater than 0.95 for all fittings (except methanol as mobile phase for C4 column). For the standard C18 column, the molar volume changes were calculated to be around －110 mL/mol and －128 mL/mol for acetonitrile and methanol respectively. For the C4 column, the values were －116 mL/mol and －56 mL/mol. For the low density C18 column, the volume change was around －168 mL/mol and －163 mL/mol for acetonitrile and methanol, respectively. In addition, the quadratic equation was also used to fit the plot of lnk' versus pressure with R2 values greater than 0.99 for all fittings. Results indicate that the molar volume change of lysozyme was decreased as the pressure increased. Moreover, the fraction of the lysozyme eluted under high pressure (300 bar) was collected and reinjected into the column at low pressure (20~30 bar). Results show that the pressure-induced change of the chromatographic behavior of lysozyme is reversible.