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Alzaabi, Adhari Abdullah. "Identification and Characterization of Serum Biomarkers Associated with Breast Cancer Progression." BYU ScholarsArchive, 2016. https://scholarsarchive.byu.edu/etd/6452.
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Despite the recognized advances in the treatment of breast cancer, it still accounts for 15% of all cancer-related deaths. 90% of breast cancer deaths are due to unpredicted metastasis. There is neither successful treatment for metastatic patients nor a specific test to predict or detect secondary lesions. Patients with primary tumor will be either over-treated with cytotoxic side effects or under-treated and risk recurrence. This necessitates the need for personalized treatment, which is hard to offer for such heterogeneous disease. Obstacles in treating breast cancer metastasis are mainly due to the gaps exist in the understanding of the molecular mechanism of metastasis. The linear model of metastasis is supported by several observations that reflect an early crosstalk between the primary and secondary tumor, which in turn makes the secondary microenvironment fertile for the growth of disseminated cells. This communication occurs through circulation and utilizes molecules which have not been identified to date. Identifying such molecules may help in detecting initial stages of tumor colonization and predict the target organ of metastasis. Furthermore, these molecules may help to provide a personalized therapy that aims to tailor treatment according to the biology of the individual tumor. Advances in proteomics allows for more reproducible and sensitive biomarker discovery. Proteomic biomarkers are often more translatable to the clinic compared to biomarkers identified using other omics approaches. Further, protein biomarkers can be found in biological fluids making them a non-invasive way to treat or investigate cancer patients. We present in this manuscript our study of the use of a proteomic approach on blood serum samples of metastatic and non-metastatic patients using LC-MS/MS quantitative analysis machine to identify molecules that could be associated with different stages of breast cancer metastasis. We focused on the deferential expression of low molecular weight biomolecules known to reflect disease-specific signatures. We manually analyzed 2500 individual small biomolecules in each serum sample of total of 51 samples. Comparisons between different sample types (from stage I and III Breast Cancer patients in this case) allows for the detection of unique short peptide biomarkers present in one sample type. We built a multi-biomarker model with more sensitivity and specificity to identify the stage of the tumor and applied them on blinded set of samples to validate prediction power. We hope that our study will provide insights for future work on the collection, analysis, and understanding of role of molecules in metastatic breast cancer.
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Parker, Todd Avery. "Elucidating the immunoactivity of a goat serum peptide." Diss., Mississippi State : Mississippi State University, 2002. http://library.msstate.edu/etd/show.asp?etd=etd-03072002-141009.
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Nash, Ian Alun. "The development of solid phase strategies and methodologies : the synthesis of polyamine toxins and peptide nucleic acids." Thesis, University of Nottingham, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.321373.
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Ali, Stuart Alvaro. "Transferrin trojan horses : a novel approach for drug delivery?" Thesis, Brunel University, 1999. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.285047.
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Huber, Alexander Domenico. "Purification of Phage-Displayed HSA-Specific Peptide for Biosensor Production." Youngstown State University / OhioLINK, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1559730074086289.
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Kenney, Floyd E. "Biosensor Production By Conjugation Of HSA-Specific Peptide To Functionalized Nanotube Fiber." Youngstown State University / OhioLINK, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1525360589515967.
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Masson, Catherine. "Purification et mode d'action d'un peptide sérique hyperglycémiant induit par la température." Nancy 1, 1990. http://www.theses.fr/1990NAN10294.
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Des travaux antérieurs ont montré qu'une hyperthermie localisée provoquait chez le rat une augmentation de la glycémie de 30 à 40%. De plus, l'injection de sérum de rat chauffé à 44#OC pendant 15 minutes à des rats normaux, entraînait également une hyperglycémie. Cet effet est lié à la présence d'un peptide dans le sérum chauffé, qui est appelée TIF (température induced factor). Dans ce travail, nous décrivons la purification de ce peptide en 4 étapes chromatographiques associant la filtration sur gel et la chromatographie liquide haute performance. Nous présentons ses propriétés physicochimiques et sa composition en acides aminés. Nous étudions les effets du sérum chauffé et du TIF sur le métabolisme glucidique du rat sain, par dosages de paramètres métaboliques (lactate, pyruvate, corps cétonique, acides aminés glucoformateurs) et hormonaux (insuline, glucagon) et étude de la cinétique de captation du glucose marqué au tritium. Une étude de l'effet hyperglycémiant, de la survie et de la taille tumorale est réalisée chez des rats porteurs de rhabdomyosarcome, et traités par injection de sérum chauffé
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Puckett, Nathan. "Effects of Binding Affinity between Bovine Serum Albumin and Platinum Drugs." TopSCHOLAR®, 2017. http://digitalcommons.wku.edu/theses/1977.
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Platinum complex drugs such as cisplatin have been used as highly successful chemotherapy drugs since the 1970s. We are interested in how the ligands attached to cisplatin analogs influences their reactivity with biologically relevant targets along with time and amount. For this study, reactions were conducted to determine the reactivity between different platinum compounds and the protein bovine serum albumin. Various platinum compounds with different ligands were reacted in varying amounts with albumin in ammonium acetate buffer for either 1 hour, 4 hours, or 24 hours. Each reaction was quenched after the designated reaction time by dialysis and the platinum bound to the protein was determined by use of ICP. LC-MS was used to find exact peptide residues platinum complexes prefer to bind with but was found to be ineffective. Results show that time has a more significant affect on binding over amount of platinum present. In respect to changing the leaving or carrier ligands on the platinum complex, these changes on the complex did not affect binding significantly with bovine serum albumin. Triamine platinum complexes also seem to bind significantly more than diamine platinum complexes along with anionic form platinum complexes binding significantly better than the cationic form platinum complexes.
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Mitchell, Emma. "Serum calcitonin gene-related peptide concentrations in the horse and their relationship to the Systemic Inflammatory response." Thesis, Virginia Tech, 2006. http://hdl.handle.net/10919/33834.
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Systemic inflammation is a leading cause of mortality and morbidity in both human and equine intensive care patients. This systemic inflammatory response may be due to insult from bacterial, viral, fungal or parasitic invasion or from trauma or hypoxemia. Local and systemic release of a wide variety of endogenous pro-inflammatory mediators results in activation of the innate immune system in order to resolve the insult. In sepsis this initial appropriate host response becomes amplified and deregulated leading to refractory hypotension and multiple organ dysfunction. The exact incidence of sepsis (SIRS due to bacterial infection) has not been reported in the equine literature (Roy 2004). Since early recognition and treatment of sepsis are associated with improved outcome the search for markers to accurately predict presence of sepsis and likelihood of survival continues. The serum concentration of both procalcitonin and its related molecule CGRP have been documented to increase in humans with SIRS, yet no literature exists as to the production or role of CGRP in equine patients with SIRS.
This study showed that equine CGRP was produced in detectable quantities by healthy adult horses and neonatal foals less than two weeks of age using a rat á-CGRP ELISA. The low percentage recovery of CGRP from samples and the high lower limit of detection for the assay prevented establishment of a normal concentration range of CGRP in healthy horses. In both adult horses and foals with documented SIRS, CGRP concentrations were significantly increased at time of presentation to the hospital (p<0.0002, p<0.003 respectively). A trend towards increased serum CGRP concentration was present in anaesethized horses exposed to endotoxin, but this was not statistically significant (p< 0.067).Master of Science
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Pham, Errek Manh Trung. "Producing A Peptide For Use In A Blood Biosensor For Injury Detection." Youngstown State University / OhioLINK, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1607519672342672.
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Rees, William D. "Identifying Peptides that Bind to Human Serum Albumin Using Phage Display for the Development of Sensors that Detect Injury in Military Personnel." Youngstown State University / OhioLINK, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1470149344.
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Tummala, Manorama. "Surfactant-Aided Matrix Assisted Laser Desorption/Ionization Mass Spectrometry (SA-MALDI MS)." University of Cincinnati / OhioLINK, 2004. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1100672049.
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Tsourapas, George. "Surface engineering of poly(_D_L-lactic acid) to create polymeric supports that inhibit serum protein interactions and promote peptide interactions." Thesis, University of Nottingham, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.428948.
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Neethling, Elmari. "Point-of-care ultrasound abnormalities in late onset severe preeclampsia: prevalence and association with serum albumin and brain natriuretic peptide." Master's thesis, University of Cape Town, 2018. http://hdl.handle.net/11427/29837.
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Abstract Background: Pilot studies applying point-of-care ultrasound (POCUS) in preeclampsia indicate the presence of pulmonary interstitial edema, cerebral edema, and cardiac dysfunction. Laboratory markers of oncotic pressure (albumin) and cardiac dysfunction (brain natriuretic peptide [BNP]) may be abnormal, but the clinical application remains unclear. We investigated the prevalence of pulmonary interstitial syndrome (PIS), cardiac dysfunction, and increased optic nerve sheath diameter (ONSD) in late onset preeclampsia with severe features. The primary aim was to examine the association between PIS or ONSD and maternal serum albumin level. The secondary aims were to explore the association between cardiac dysfunction and PIS, ONSD, BNP, and serum albumin level, and between POCUS-derived parameters and a suspicious or pathological cardiotocograph (CTG). Methods: Ninety-five women were enrolled in this prospective observational cohort study. A POCUS examination of lungs, heart and ONSD was performed. PIS was defined as a bilateral B-line pattern on lung US, and diastolic dysfunction according to an algorithm of the American Society of Echocardiography. ONSD > 5.8 mm was interpreted as compatible with raised intracranial pressure (> 20 mmHg). Serum BNP and albumin levels were also measured. Results: PIS, diastolic-, systolic dysfunction, and raised left ventricular end-diastolic pressure (LVEDP) were present in 23 (24%,) 31 (33%), 9 (10%), and 20 (25%) women respectively. ONSD was increased in 27 (28%) women. Concerning the primary outcome, there was no association between albumin level and PIS (p = 0.4) or ONSD (p=0.63). With respect to secondary outcomes, there was no association between albumin level and systolic dysfunction (p = 0.21) or raised LVEDP (p = 0.44). PIS was associated with diastolic dysfunction (p = 0.02), and raised LVEDP (p = 0.009, negative predictive value 85%). BNP level was associated with systolic (p < 0.001)- and diastolic dysfunction (p = 0.003) and LVEDP (p = 0.007). No association was found between POCUS abnormalities and a suspicious/pathological CTG (p = 0.07). Conclusion: PIS, diastolic dysfunction and increased ONSD were common in preeclampsia with severe features. Cardiac ultrasound abnormalities may be more useful than albumin levels in predicting PIS. The absence of PIS may exclude raised LVEDP. The further clinical relevance of PIS and raised ONSD remains to be established. BNP level was associated with cardiac ultrasound abnormalities. Although this study was not designed to directly influence clinical management, the findings suggest that POCUS may serve as a useful adjunct to clinical examination for the obstetric anesthesiologist managing these complex patients.
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Faradj, Lana. "Improving Caco-2 cell permeability assay using phospholipid covered silica beads." Thesis, Uppsala universitet, Institutionen för farmaci, 2021. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-432865.
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The Caco-2 cell assay is widely used for in vitro permeability measurements. However, a draw back with the assay that this study will focus on improving, is compound adsorption to the plastic material. Lipophilic compounds such as Cyclosporin A and Peptide J, that will be used in this study, tend to bind to the plastic material in the assay. This can result in poor recovery and misleading permeability predictions. Bovine serum albumin (BSA) is an alternative used today to prevent this but is not always successful. The aim of this study is therefore to improve the Caco-2 permeability assay by adding phospholipid covered silica beads (PLB) to the basolateral chamber. The role of the PLB is to bind the compound of interest and decrease the amount of compound bound to the plastic material and thus better predict the permeability of the compound of interest. The PLB was produced using phosphatidylcholine and silica beads. Caco-2 cells were seeded and maintained for 21-29 days ahead of the experiment. PLB concentration of 20, 60 and 100 mg/ml were prepared. Samples were analyzed with HPLC-MSMS. The results showed that with increasing PLB concentration we had a significantly decrease in non-specific plastic binding resulting in reliable permeability predictions, concluding that the hypothesis was correct.
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Greiff, Victor [Verfasser], Michal [Akademischer Betreuer] Or-Guil, Edda [Akademischer Betreuer] Klipp, and Birgit [Akademischer Betreuer] Sawitzki. "Exploring the genesis and specificity of serum antibody binding : mathematical modeling and data analysis of antibody-peptide reactivity data / Victor Greiff. Gutachter: Michal Or-Guil ; Edda Klipp ; Birgit Sawitzki." Berlin : Humboldt Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2012. http://d-nb.info/1030313598/34.
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Greiff, Victor [Verfasser], Michal Akademischer Betreuer] Or-Guil, Edda [Akademischer Betreuer] [Klipp, and Birgit [Akademischer Betreuer] Sawitzki. "Exploring the genesis and specificity of serum antibody binding : mathematical modeling and data analysis of antibody-peptide reactivity data / Victor Greiff. Gutachter: Michal Or-Guil ; Edda Klipp ; Birgit Sawitzki." Berlin : Humboldt Universität zu Berlin, Mathematisch-Naturwissenschaftliche Fakultät I, 2012. http://nbn-resolving.de/urn:nbn:de:kobv:11-100206822.
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EGARD, THIERRY. "Recherche d'une reactivite vis-a-vis d'un peptide correspondant a la sequence 69-84 de hla-b*2705 dans le serum de patients atteints d'une spondylarthrite ankylosante ou d'une arthrite reactionnelle." Lille 2, 1989. http://www.theses.fr/1989LIL2M442.
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Kubeil, Manja. "Design und Synthese von mehrfunktionalen Cyclamliganden zur Entwicklung von stabilen radioaktiven Kupferkomplexen für Diagnostik und Therapie." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2014. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-142867.
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Die Entwicklung von makrocyclischen Chelatoren, die mit Kupferionen thermodynamisch stabile und kinetisch inerte Komplexe bilden, ist in den letzten Jahren zunehmend in den Fokus der Forschung gerückt. Das ergibt sich insbesondere aus der Möglichkeit, Radiokupfernuklide aufgrund günstiger kernphysikalischer Eigenschaften sowohl für diagnostische (64Cu) als auch therapeutische (67Cu) Anwendungen einzusetzen.
Hervorzuheben ist der Azamakrocyclus Cyclam (1,4,8,11-Tetraazacyclodecan), da dieser mit Kupfer(II)-Ionen Komplexe hoher thermodynamischer Stabilität bildet. Zudem weist der Chelator weitere Funktionalisierungsmöglichkeiten auf, um in Hinblick auf eine nuklearmedizinische Anwendung pharmakologisch relevante Moleküle wie beispielsweise Peptide oder Proteine (Antikörper oder Antikörperfragmente) kovalent zu binden und damit eine spezifische Anreicherung im Tumorgewebe zu ermöglichen.
Allerdings erfordert das Maßschneidern der Eigenschaften von neuen bifunktionellen Chelatoren Kenntnisse über den Einfluss der Substituten auf die Stabilität der gebildeten Kupfer(II)-Komplexe. Die thermodynamische Stabilität lässt keine Aussagen über das Verhalten in vivo zu. Die Ursache für die kinetische Labilität in Säugetieren ist noch nicht vollständig verstanden, wird aber u. a. auf kupferbindende Enzyme bzw. Proteine zurückgeführt. Die Bioverteilung der Radiokupferkomplexe wird aber auch von weiteren Parameter, wie Hydrophilie, Ladung und Polarisierbarkeit beeinflusst.
Mit dieser Arbeit wurde ein wesentlicher Beitrag zur Entwicklung von 64Cu Chelaten auf Basis von Cyclam-Propionsäure-Liganden geleistet. Diese Stoffklasse ist bisher synthetisch wenig erschlossen und Radiokupfer-markierte Komplexe zudem bisher gar nicht beschrieben. Daher ist es von besonderem Interesse die kinetische Stabilität Radiokupfer-markierter Cyclam-Propionsäure-Derivate zu untersuchen und mit einer Reihe bekannter Radiokupfer-markierter Cyclam-Essigsäure-Komplexe zu vergleichen.
Es wurden vier N-funktionalisierte Cyclam-Derivate 13, 14b, 15 und 16, die eine unterschiedliche Anzahl an Propionsäuregruppen tragen, erfolgreich synthetisiert und in sehr hoher Reinheit (>99%) isoliert. Besonders hervorzuheben ist die erstmalige Synthese des trans-N,N´´-funktionalisierten Cyclam-Propionsäure-Derivates 14b in hoher Ausbeute (gesamt = 32%). Von den Verbindungen 13, 14b, 15 und 16 sind entsprechende Kupfer(II)-Komplexe hergestellt worden. Zur Aufklärung relevanter Fragestellungen bezüglich der chemischen und geometrischen Eigenschaften sind verschiedene spektroskopische Methoden (Röntgeneinkristallstrukturanalyse, IR, UV/VIS, Elektronenspinresonanz-Spektroskopie) anhand von den isolierten Kupfer(II)-Komplexen CuII-13, CuII-14b und CuII-16 herangezogen worden.
Die röntgenkristallografischen Strukturaufklärungen der Komplexe CuII-14b und CuII-16 weisen eine verzerrt quadratisch-pyramidale Koordinationsgeometrie auf. Das Ligandenfeld wird innerhalb der Stickstoffebene mit steigendem Substitutionsgrad schwächer. Das wurde auch durch ESR-Messungen bestätigt. Nachweislich verursachen die zusätzlichen funktionellen Gruppen eine kleinere Ligandenfeldaufspaltung. Weiterhin nahm die kinetische Stabilität unter stark sauren Bedingungen mit steigendem Substitutionsgrad ab. Der Vergleich mit den bekannten oktaedrischen Kupfer(II)-Cyclam-Essigsäure-Komplexen zeigt, dass die quadratisch-pyramidalen Kupfer(II)-Cyclam-Propionsäure-Derivate unter stark sauren Bedingungen schneller dissoziieren. Als Ursache können die unterschiedlichen Konfigurationen diskutiert werden, da bei 4N+2-Geometrien die thermodynamisch bevorzugte trans III-Konfigurationen gebildet wird.
Radiochemische Untersuchungen zur Bewertung der kinetischen Stabilität in vitro und in vivo sind mit den 64Cu-markierten Liganden 13, 14b, 15 und 16 durchgeführt worden. Hierfür ist ein In-vitro-Stabilitätstest basierend auf dem kupferbindenden Enzym Superoxid-Dismutase (SOD) bzw. humanem Serum für radiomarkierte Verbindungen entwickelt worden. In humanem Serum ist Albumin (~66 kDa) in sehr hoher Konzentration enthalten und eines der wichtigsten Transportproteine für extrazelluläre Kupfer-Ionen. Aufgrund seiner Abundanz im Blutplasma ist im Serum-Assay jeweils nur eine stark ausgeprägte Bande bei ca. 66 kDa detektiert worden. Dieser etablierte In-vitro-Stabilitätstest beruht im Gegensatz zu anderen herkömmlichen Analysemethoden (Radio-HPLC oder Radio-DC) auf dem Prinzip der Gelelektrophorese. Von großem Vorteil ist, dass mehrere Proben simultan untersucht werden können und die Ergebnisse zuverlässig und reproduzierbar sind.
Die In-vitro-Ergebnisse zeigen einen ähnlichen Trend wie bei der säure-assoziierten Dissoziation, wobei die höchste Stabilität bemerkenswerterweise bei dem [64Cu]Cu-14b Komplex bestimmt wurde. Allerdings beruht der Mechanismus hier nicht auf einer Dissoziation sondern auf einer Transchelatisierung. Die Ergebnisse der Bioverteilungen in Wistar-Ratten korrelieren mit den In-vitro-Studien in humanem Serum. Der Komplex [64Cu]Cu 14b zeigte sowohl eine schnelle renale Blut-Clearence als auch eine sehr geringe Anreicherung in der Leber und stellt damit eine Alternative zu den kommerziell erwerblichen Liganden dar. Als geeignete Chelatoren bieten Cyclam-Monopropionsäure 13 und Cyclam-Dipropionsäure 14b die Möglichkeit, Radiokupfernuklide stabil zu binden und erlauben die mehrfache Einführung von EGFR-spezifischen Peptiden an das Grundgerüst. Als Grundgerüst wurde der Ligand 13 ausgewählt.
Durch die Multifunktionalisierung sollen höhere Affinitäten zum Rezeptor und verbesserte metabolische Stabilitäten hervorgerufen werden. Für diese Verbindung liegen erste vielversprechende Ergebnisse vor, wobei hohe Affinitäten zu zwei EGFR-positiven Zelllinien bestimmt wurden.
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Olszewski, Amy L. "Synthesis, Biological Functionalization, and Integration ofCarbon Nanotubes for Bio-Sensing Textiles." Youngstown State University / OhioLINK, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1369854838.
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Schamber, Christine [Verfasser], Piotr [Akademischer Betreuer] Lewczuk, and Johannes [Akademischer Betreuer] Kornhuber. "Einflussfaktoren auf die Plasmakonzentration der Amyloid-β-Peptide Aβ1-40, Aβx-40, Aβ1-42 und Aβx-42 in einem Probandenkollektiv kognitiv unbeeinträchtigter Erwachsener - Eine Untersuchung der Faktoren Body Mass Index, Blutdruck, Glukose-, Triglyzerid- und Kreatininkonzentration im Serum sowie biographischer und familienanamnestischer Daten / Christine Schamber. Gutachter: Piotr Lewczuk ; Johannes Kornhuber." Erlangen : Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), 2015. http://d-nb.info/107616661X/34.
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Tengel, Tobias. "Studies of protein structure, dynamics and protein-ligand interactions using NMR spectroscopy." Doctoral thesis, Umeå : Univ, 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-1472.
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Jacksén, Johan. "Improved techniques for CE and MALDI-MS including microfluidic hyphenations foranalysis of biomolecules." Doctoral thesis, KTH, Analytisk kemi, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-27342.
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In this thesis, improved techniques for biomolecule analysis using capillary electrophoresis (CE) and matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) and hyphenations between those have been presented.A pre-concentration method which is possible to apply in both techniques, has also been investigated. In this work the off-line MS mode has been used either in the form of fractionation (Paper I) or by incorporating the MALDI target in the CE separation system (Paper II).In Paper I, a protocol for CE-MALDI analysis of cyanogen bromide digested bacteriorhodopsin (BR) peptides as model integral membrane protein peptides were established. Also, an improved protocol for partially automated manufacturing of a concentration MALDI-target plate is presented. The design of the targets was suitable for the fractions from the CE. A novel technique for the integration of CE to MALDI-MS using a closed-open-closed system is presented in Paper II, where the open part is a micro canal functioning as a MALDI target window. A protein separation was obtained and detected with MALDI-MS analysis in the micro canal. A method has been developed for detection of monosaccharides originating from hydrolysis of a single wood fiber performed in a micro channel, with an incorporated electromigration pre-concentration step preceding CE analysis in Paper III. The pre-concentration showed to be highly complex due to the fact that several parameters are included that affecting each other. In Paper IV a protocol using enzymatic digestion, MALDI-TOF-MS and CE with laser induced fluorescence (LIF) detection for the investigation of the degree of substitution of fluorescein isothiocyanate (FITC) to bovine serum albumin (BSA), as a contact allergen model system for protein-hapten binding in the skin, is presented. The intention of a further CE-MALDI hyphenation has been considered during the work. In Paper V 2,6-dihydroxyacetophenone (DHAP) was investigated, showing promising MALDI-MS matrix properties for hydrophobic proteins and peptides. 2,5-dihydroxybenzoic acid (DHB) was undoubtedly the better matrix for the hydrophilic proteins, but its performance for the larger and hydrophobic peptides was not optimal. Consequently, DHAP can be used as a compliment matrix for improved analysis of hydrophobic analytes.QC 20101214
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Nelson, Geoffrey Winston. "Surface characterization and functional properties of carbon-based materials." Thesis, University of Oxford, 2012. http://ora.ox.ac.uk/objects/uuid:f22b95ce-65f3-4d9e-ac3d-a88f6e142c1a.
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Carbon-based materials are poised to be an important class of 21st century materials, for bio-medical, bio-electronic, and bio-sensing applications. Diamond and polymers are two examples of carbon-based materials of high interest to the bio-materials community. Diamond, in its conductive form, can be used as an electrochemical bio-sensor, whilst its nanoparticle form is considered a non-inflammatory platform to deliver drugs or to grow neuronal cells. Polymers, especially when chemically modified, have been used extensively in biological environments, from anti-microbial use to drug delivery. The large-scale use of either material for biological use is limited by two factors: ease of chemical modification and the paucity of knowledge of their surface chemistry in aqueous media. This thesis addresses aspects of both these issues. The first study reported is an in situ study of the adsorption dynamics of an exemplar globular protein (bovine serum albumin, BSA) on nanodiamond using the relatively novel quartz crystal microbalance with dissipation (QCM-D) technique. For the first time, QCM-D enabled the detailed study of protein dynamics (i.e. kinetics, viscoelastic properties, overlayer structure, etc.) onto nanodiamond thin films having various surface chemistry and roughness. The dynamics of protein adsorption is found to be sensitive to surface chemistry at all stages of adsorption, but it is only sensitive to surface roughness during initial adsorption phases. Our understanding of the nanodiamond-biology interface is enhanced by this study, and it suggests that QCM-D is useful for the study of the surface chemistry of nanoparticle forms of inorganic materials. A second study concerns a novel surface functionalization scheme, based on carbene and azo-coupling chemistry, which has been recently introduced as a practical, facile method for modifying the surfaces of polymers. Using modern surface characterization techniques, it is demonstrated that a chemical linker can be attached to polystyrene surfaces using carbene-based chemistry, and that further chemical functionality can be added to this chemical linker via an azo-coupling reaction. In situ studies of protein dynamics at these interfaces were conducted using QCM-D, thus enabling a link between specific protein behaviour and the polymer surface chemical termination chemistry to be made. A third area of study of investigates the use of diamond electrodes as a bio-sensor for dopamine under physiological conditions. For these conditions, ascorbic acid interferes with the dopamine oxidation signal, in ways that render the two signals irresolvable. Various modifications are used in attempts to reduce this interference, including: small and large cathodic treatments, grafting of electro-active polymers, addition of carbon nanotubes, and hydrogen plasma treatment. Those modifications leading to the hydrogen-termination of diamond are shown to work the best. Notably, hydrogen plasma treatment effects the complete electrochemical separation of dopamine and ascorbic acid at a diamond electrode. This is the first time this has been accomplished without adding non-diamond materials to the diamond electrode surface.
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Such, Sanmartin Gerard. "Assessing human growth hormone variants to determine their potential relevance in anti-doping and clinical analysis." Doctoral thesis, Universitat Pompeu Fabra, 2010. http://hdl.handle.net/10803/7198.
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Human growth hormone (GH) participates in human longitudinal growth, lipid and carbohydrate metabolism, and comprises a remarkably number of proteins with similar sequences, generated either genetically or post-translationally, that in some cases show clearly differentiated biological activities. Current methods employed for its quantification are mainly based on a specific immunodetection of the most concentrated GH variants in blood circulation. However, it is not clear neither which variants are recognised in each case, nor which is the real concentration of some of these variants. Probably related, present immunoassays show a disparity of results between them that difficults the comparison of data from different assays, with direct consequences in the clinical field. Within a doping context, the illegal administration of recombinant GH constitutes a complex challenge, given the fact that the pharmaceutical variant and the native 22 kDa GH variant do not show any structural difference that allows a direct detection. However, the administration of the pharmaceutical inhibits the natural production of the hormone, resulting in modifications between the relative concentration of some of these variants. In this case, which variants are detected is of utmost importance, since these constitute the base of this anti-GH doping method. Here, the relevance of some GH variants is addressed, including their generation, characterisation, analysis through specific antibodies and their detection on biological samples.L'hormona de creixement humana (GH) participa en el creixement post-longitudinal i en el metabolisme de lípids i carbohidrats, i comprèn un extraordinari nombre de proteïnes de seqüències similars, generades tant genèticament com posttranslacional, que en alguns casos mostren activitats biològiques clarament diferenciades. Els mètodes actuals emprats per la seva quantificació es basen principalment en una immunodetecció específica de les variants de GH més concentrades en circulació sanguínia. Tanmateix, no resta clar quines variants es reconeixen en cada cas, ni quina és la concentració real d'algunes d'aquestes variants. Possiblement relacionat, els immunoassaigs actualment utilitzats mostren una disparitat de resultats que dificulten la comparació de dades d'assaigs diferents, amb conseqüències directes en el camp clínic. Dins d'un context de dopatge, l'administració il·legal de GH recombinant constitueix un desafiament complex, donat el fet que la variant farmacèutica i la variant de GH nativa de 22 kDa no mostren cap diferència que permeti una detecció directa. No obstant, l'administració del medicament farmacèutic inhibeix la producció natural de la hormona, derivant en canvis entre la concentració relativa d'algunes d'aquestes variants. En aquest cas, és de màxima importància quines variants són detectades, ja que això constitueix la base d'aquest mètode d'antidopatge de GH. Aquí, s'estudia la rellevància d'algunes variants de GH, incloent-hi la seva generació, caracterització, anàlisi via anticossos específics i la seva detecció en mostres biològiques.
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Chen, Yu-hsien, and 陳玉賢. "Prolonging serum half-lives of peptide drugs by site-specific chemical modifications." Thesis, 2001. http://ndltd.ncl.edu.tw/handle/60085931576987265045.
Full textAbstract:
碩士國立清華大學
生命科學系
89
Abstract Prolonging serum half-lives of protein/peptide drugs in vivo is important in drug delivery systems. Site-specific chemical modification is one of the approaches. By choosing a suitable site of proteins and controlling the reaction condition, we can obtain the homogeneous product without affecting the activity of proteins. Because of the high reactivity of thiol group, cysteine is a good choice to react with some sulfhydryl-specific modifiers. Here, we mutated a phenylalanine39 of ACTH to cysteine (ACTH-F39C). The free sulfhydryl group of cysteine then reacted with two kinds of chemical modifiers─iodoacetamide derivatives with long carbon chains and methoxy-polyethylene glycol (m-PEG) maleimide derivatives. After lipophilic modification, the in vitro biological activities of these adducts were not disturbed compared to ACTH wild type. In ELISA and flow cytometry experiments, F39C-12CH3 is associated more strongly with plasma membranes and serum albumins than the unmodified one. We injected unmodified ACTH and the lipophilic adducts to mice by intramuscular injection. The maintaining percentage in mice serum of F39C-12CH3 is 3-folds higher than F39C after 6 hours. In PEG modification, the in vitro activities of F39C-PEG20K and F39C-PEG40K were slightly decreased. After 24 hours，the remaining percentage of F39C-PEG20K and F39C-PEG40K in mice serum is 6-folds higher than F39C. We concluded that chemical modifications to change the lipophilicity or molecular size of proteins/peptides could prolong the serum half-life in vivo.
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Bissegger, Sonja. "Development of a Supplement for CHO Cell Culture Serum-free Media by the Fractionation of Peptide mixtures using Nanofiltration." Thesis, 2009. http://hdl.handle.net/10012/4868.
Full textAbstract:
The objective of this work was the investigation of nanofiltration as a potential avenue to fractionate protein hydrolysates and produce protein hydrolysate fractions with stimulating bioactivity for the development of a supplement for a serum-free media.
Mammalian cell culture is widely used for the production of therapeutic proteins such as antibodies, interleukins, and vaccines because of the ability of mammalian cells to glycosylate proteins. A complex media with the addition of serum is often required to meet the requirements of the cells. Although serum is a supplement that provides different proteins such as growth factors and hormones, serum has several disadvantages such as high cost, difficulty of downstream processing due to its high protein content and the possibility of microbiological contamination. Protein hydrolysates from plant, animal, or yeast cells contain a complex mixture of peptides and amino acids and have been shown to enhance growth of certain mammalian cell lines cultured in serum-free media.
To fractionate peptide mixtures, nanofiltration was investigated in this study. Nanofiltration is a pressure driven membrane separation process based on size and charge. The investigation of pH and NaCl on the filtration performance for two different nanofiltration membranes (HL membrane and G-10 membrane) was achieved using a 24 factorial design. The total peptide concentration, the antioxidant activity, and organic and inorganic content were analyzed in the permeate and retentate fraction. The fractions were also tested for their enhanced growth ability and the specific -interferon productivity with CHO cells. Furthermore the retentate and permeate fractions were analyzed by reversed phase-HPLC to characterize the peptide and free amino acid distribution profile.
Through the factorial design, the membrane type was shown to have a significant effect on the filtration performance for both yeast extract and Primatone. A significant difference, but similar for both feed sources, was observed for the total peptide transmission with around 10% for the HL membrane and around 30% for the G-10 membrane. The average permeate flux was significantly lower for the G-10 membrane although the G-10 membrane is a loose nanofiltration membrane with a reported 2500 Da MWCO compared to the HL membrane with a reported 300-500 Da MWCO. The total peptide transmission, organic and inorganic content of the fractions for the two feed sources and membrane type were affected differently according to pH and NaCl addition. These results indicate that the two feed sources are of different composition and that nanofiltration is a possible method to fractionate peptides.
The bioactivity of the nanofiltration fractions was tested as a nutrient additive to a serum-free media in CHO cells. It was shown that the productivity is not always related to the cell density, as the highest overall specific interferon productivity was achieved for low cell density similar to the hydrolysate free negative control. Furthermore, the retentate fraction of yeast extract separated with the G-10 membrane at a pH of 8 resulted in the highest cell density. According to these results, nanofiltration is a promising method for the enrichment of protein hydrolysates as a supplement for serum in cell culture.
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Kuo, Chia-Tzu, and 郭佳詞. "Development of amine modified graphene oxide based peptide SPR aptasensor for human chorionic gonadotropin in clinical serum detection." Thesis, 2017. http://ndltd.ncl.edu.tw/handle/ex4vne.
Full textAbstract:
碩士國立臺灣師範大學
光電科技研究所
105
In this work we present a facile processes amine modified graphene oxide (GO-NH2) as an ultra-sensitive surface plasmon resonance (SPR) probe grafting layer for non-immunization sensing. The GO-NH2 based SPR biosensor was applied in detection of human Chorionic Gonadotropin (hCG), relevant to pregnant disease. Such as premature birth, ectopic pregnancy, and Down's syndrome. It’s also relevant to some cancer about gonad, like ovarian cancer, breast cancer, or testicular cancer. The concentration of hCG in blood plasma of healthy and non-pregnant is around 0.45 nM. Replacing antibody by non-immunization peptide got an opportunity of preserve the probe in relative higher temperature and much lower cost than antibodies. In order to diagnose the patient in the early state and benefit for commercial situation, the characteristic of high resolution GO-NH2 modified bio-chip is a novel and advantage technique. The first part of two-step modified approach is substitute the hydroxyl with chloride on GO. The chloride functional graphene oxide (GO-Cl) is obtained through the reaction of GO and thionyl chloride (SOCl2). After that, we utilize ammonia water as nitrogen precursor, and the chloride groups are replaced by amino groups. The primary of facial amine group is identified by X-ray photoelectron spectroscopy (XPS). The ratio of nitrogen and carbon in our experimental parameter is increase to 5.7%, and the ratio of oxygen is changed from 27% to 17%. In the Fourier transform infrared spectroscopy (FTIR) experiment. The absorbed peak of 1064 〖cm〗^(-1), 1579 〖cm〗^(-1), and 3361 〖cm〗^(-1) that absorbed by N are found. The simulation of complex refractive index and the impedance of GO-NH2 is 1.55 + C_1 λ/n i and 89.16 Ω. The recombinant protein experiment shows that GO-NH2 sensor-chip is 2.45 times about the affinity of peptide. Comparing in non-immunization diagnostic, the response at 2 nM is 2.68 times greater, and the slope of GO-NH2 linear regression is 1.5 times higher than commercial chip. Last, the clinical blood serum experiment shows high linear regression coefficients.
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Wu, Hsiu-Lin, and 吳秀琳. "The investigation of autoantibody isotypes against novel modified acrolein C4 complement peptide adduct in serum from rheumatoid arthritis." Thesis, 2018. http://ndltd.ncl.edu.tw/handle/68z6v5.
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Huang, Ching-Wen, and 黃靖雯. "The investigation of autoantibody isotypes against novel acetylation modified peptide adducts in serum from Taiwanese females with Systemic Lupus Erythematosus." Thesis, 2018. http://ndltd.ncl.edu.tw/handle/x8f5aj.
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Chen, Shu-Yi, and 陳淑怡. "The Investigation of malondialdehyde modification autoantibody isotype against novel α-2-macroglobulin peptide in serum from Taiwanese women with Systemic Lupus Erythematosus." Thesis, 2018. http://ndltd.ncl.edu.tw/handle/e87349.
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Hsueh, Tung, and 薛彤. "Clinical Application of Single and Serial Measurements of Serum N-terminal pro- brain Natriuretic Peptide Concentration in Dogs with Degenerative Mitral Valve Disease." Thesis, 2017. http://ndltd.ncl.edu.tw/handle/01274141924581092543.
Full textAbstract:
碩士國立中興大學
獸醫學系暨研究所
105
The N-terminal pro-brain natriuretic peptide (NT-proBNP) is released from stretched cardiomyocytes and its circulating concentration is positively associated with heart disease severity. The purpose of this study was to evaluate the correlation between NT-proBNP and various clinical assessments, and to estimate the prognostic value of serial measurement of serum concentration of NT-proBNP in dogs with congestive heart failure (CHF) secondary to degenerative mitral valve disease (DMVD). Eighty client-owned dogs were enrolled and classified by American College of Veterinary Internal Medicine guideline, and they were also allocated into groups by cardiac remodeling and CHF. Thirty-four dogs with CHF further underwent serial measurements of NT-proBNP. The mortality rate between sub-categorical groups were compared. The results showed that NT-proBNP concentration had an increasing trend from stage A to stage B2, and was significantly higher in dogs with cardiac remodeling (p<.001) and dogs with CHF (p<.001). Moreover, NT-proBNP was significantly and positively correlated with clinical score (p<.001), radiographic score (p<.001) and several selected echocardiographic indices (all p<.001). In the survival analysis, the NT-proBNP of baseline (p=.830), first follow-up (p=.650), absolute (p=.470) and relative change (p=.376) had no significant difference between survivors and non-survivors. And the mortality rate between sub-categorical groups did not differ statistically. A possible sexual effect was observed by sex stratification. The study confirmed the clinical utility of NT-proBNP in assessing heart disease severity and in reflecting the instant clinical status in dogs with DMVD. Further study for evaluating the association among NT-proBNP trending, sex and neutered status was warrant.