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Dissertations / Theses on the topic 'Affinity'

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Published: 4 June 2021
Last updated: 2 November 2024

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1

Crawl, Lester Daniel. "Affinity-directed mobility." Diss., Connect to online resource, 2006. http://gateway.proquest.com/openurl?url_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&res_dat=xri:pqdiss&rft_dat=xri:pqdiss:3219008.

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Smith, Matthew. "Implicit affinity networks /." Diss., CLICK HERE for online access, 2007. http://contentdm.lib.byu.edu/ETD/image/etd1682.pdf.

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3

Smith, Matthew Scott. "Implicit Affinity Networks." BYU ScholarsArchive, 2007. https://scholarsarchive.byu.edu/etd/1112.

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Although they clearly exist, affinities among individuals are not all easily identified. Yet, they offer unique opportunities to discover new social networks, strengthen ties among individuals, and provide recommendations. We propose the idea of Implicit Affinity Networks (IANs) to build, visualize, and track affinities among groups of individuals. IANs are simple, interactive graphical representations that users may navigate to uncover interesting patterns. This thesis describes a system supporting the construction of IANs and evaluates it in the context of family history and online communities.
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4

Arnold, Lindsay G. "Engineering thermo-responsive affinity ligands for glycoprotein purification by affinity precipitation." Diss., Georgia Institute of Technology, 2015. http://hdl.handle.net/1853/53493.

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Effective methods for isolation and purification of glycoproteins are of increasing significance to the rapidly growing biopharmaceutical and diagnostic industry. Glycoproteins represent the majority of therapeutic proteins on the market and are effectively used to treat immune disorders, infections, cancers, and other diseases. Targeting these glycoproteins is also critical to an emerging field of glycoproteomics aimed to understand structure-function relationships of glycans. Architecturally, these glycoproteins are proteins with covalently linked oligosaccharide chains of varying monosaccharide composition. Affinity chromatography has proven to be an excellent method of glycoprotein purification at the bench scale. However, chromatography in large scale production has its drawbacks. Column fowling, flowrate limitations, and diffusional constraints collectively hinder the effectiveness of the method. An alternative proposed in this dissertation is the use of affinity precipitation as a purification technique. The three main objectives are 1) develop and produce dual-functional, thermo-responsive affinity ligands from a biological host, 2) characterize and optimize the accompanying affinity precipitation method, and 3) apply the ligand and process to relevant, unmodified glycoproteins. The design of the thermo-responsive affinity construct was comprised of two main functional domains. The binding capability was achieved by selection of small ligands with affinity to a specific monosaccharide moiety. Two different lectins, or sugar binding proteins, were used in the fusion design: a fucose binding lectin from Ralstonia solanacearum, and a sialic acid binding lectin from Vibrio cholera. The thermo-responsive functionality was obtained by use of an elastin-like peptide (ELP), which confers inverse solubility relationship properties to the fusion construct. A small library of varying ELP chain lengths were designed to find the optimal size fusion for both production and function. These dual functional ligands were cloned and expressed in the microbial host, E. coli. Furthermore, secretion of these constructs was achieved by employing the Tat secretion pathway in combination with an outer membrane lipoprotein deletion mutant with a leaky periplasm phenotype. This secretory mechanism allows for easy isolation, avoidance of inclusion bodies, and no additional protease inhibitors. After successful production, the ligands were tested to confirm that dual functionality was preserved in fusion form. Once binding conditions and precipitation properties were ascertained, the purification ability was tested on model glycoproteins. Experimentation was carried out monitoring the purification yield, purity, and retained activity of the target enzymes. High contaminant solutions, such as cell lysates, were spiked with the model glycoproteins to mimic crude protein solutions. The purification ability of the constructs in these models was observed. The method was then implemented on two relevant glycoprotein applications: 1) purification of soybean peroxidase from a crude protein extract and 2) targeting the therapeutic protein erythropoietin from albumin rich, used CHO cell media. By implementation of the fucose targeting fusion construct, the unmodified soybean peroxidase is isolated from a natural crude extract from the soybean hull, a by-product of the soybean industry. The affinity precipitation method parameters were optimized with respect to ratios, temperatures, recycle, and elution buffers to achieve successful isolation of the low abundance enzyme. Under the optimized conditions, >95% recovery yield and a purification of 22.7 fold of an active, pure product was attainable. The purification of erythropoietin led to additional experimentation with high-abundant glycoprotein solutions, as well as expansion of the affinity ligand platform. The concept of multi-lectin affinity precipitation, using the fucose and sialic acid binding lection sequentially, was introduced and tested for purification capability. An industrially relevant scheme involving isolation of the erythropoietin from used CHO cell media allowed for an achievable yield of about 60%, with a resulting albumin depletion of about 85%. In addition to development of a pair of novel thermo-responsive affinity ligands for glycoprotein purification, this dissertation provides insight on possible improvements and future directions with respect to the thermo-responsive affinity ligand platform. This unique concept employs novel lectin fusions to target valuable glycoproteins using a method avoiding the major drawbacks associated with chromatography.
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5

Campbell, Alyson Ann. "Affinity precipitation of proteases." Thesis, Heriot-Watt University, 1996. http://hdl.handle.net/10399/721.

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6

Low, Nigel Murray. "Mimicking antibody affinity maturation." Thesis, University of Cambridge, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.364567.

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7

Zourna, Kalliopi. "Smart magnetic affinity adsorbents." Thesis, University of Birmingham, 2009. http://etheses.bham.ac.uk//id/eprint/511/.

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As the focus of research on ‘adaptive/responsive’ surfaces has in recent years contributed strongly towards the design of surface materials with ‘intelligent’ or ‘smart’ behaviour, current superparamagnetic adsorbents being employed both in small and large scale operations can be surface modified and improved by gaining dual functionalities. In this work, modification of M-PVA supports with polymer brushes of dual properties has been explored for their intended use in bioseparation technology, i.e. for both selectively protein binding and enhanced temperature elution of especially difficult to elute species such as haemoglobin. Tethering of polymer brushes was achieved by employing two different ‘grafting from’ routes, i.e. cerium (IV) initiated polymerisation and Atom Transfer Polymerisation Reaction (ATRP). By identifying the optimum cerium (IV) reaction conditions, the said chemistry was further utilised to attach different polymers (thermoresponsive and affinity ligands) and their combination (thermo-affinity) at fixed positions onto M-PVA supports, either as di-block or mixed functionality polymer brushes. The configuration of introduced polymer chains as well as the haemoglobin binding characteristics of the above materials was evaluated, and their efficiency for haemoglobin and GFP desorption via sequential temperature transitions was demonstrated. Mixed polymer brushes manufactured using sequential ATRP after partial bromination of AGE activated magnetic supports were characterised and tested likewise. Protein binding and release efficiency was dependent on brush configuration (length and spacing between the graft sites of polymers), pNIPAAm content, type of affinity ligand and type of protein employed. From the above materials those with polymer chains of sufficient pNIPAAm length and at such spacing allowing their ‘free’ expansion/collapse upon temperature change (especially those grafted via cerium (IV) route) were found efficient, as brush behaviour favour enhanced desorption of difficult to elute species.
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8

Deshpande, S. "Functionalised macroporous affinity matrices." Thesis(Ph.D.), CSIR-National Chemical Laboratory, Pune, 2007. http://dspace.ncl.res.in:8080/xmlui/handle/20.500.12252/2584.

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9

Shookhoff, Alexandra. "Affinity groups commonality in diversity /." Diss., Connect to the thesis, 2006. http://hdl.handle.net/10066/606.

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10

Horstmann, Brenda Joan. "Affinity adsorption on agarose matrices." Thesis, University of Cambridge, 1989. https://www.repository.cam.ac.uk/handle/1810/250951.

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11

Gadgil, William Himanshu S. "Advances in DNA Affinity Chromatography." View the abstract Download the full-text PDF version, 2001. http://etd.utmem.edu/ABSTRACTS/2001-001-gadgil-index.htm.

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Thesis (Ph.D.)--University of Tennessee Health Science Center, 2001.
Title from title page screen (April 10, 2008). Research advisor: Harry W. Jarrett, Ph.D. Document formatted into pages (xv, 162 p. : ill.). Vita. Abstract. Includes bibliographical references (p. 152-162).
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12

Burton, Nicolas Paul. "Novel ligands for affinity chromatography." Thesis, University of Cambridge, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.359769.

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13

Santori, Fabio. "Lectin affinity chromatography of monosaccharides." Thesis, University of Bath, 2002. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.760807.

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14

Palanisamy, Uma Devi. "Affinity ligands with glycoform specificity." Thesis, University of Cambridge, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.624896.

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15

Qian, Jianing. "Affinity chromatography of camelid antibodies." Thesis, University of Cambridge, 2012. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.610171.

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16

Praig, Vera Gertraud. "Immobilised glutathione for affinity binding." Thesis, University of Cambridge, 2002. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.620379.

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Vesole, Steven Michael. "Affinity-Based Delivery of Retinoids." Case Western Reserve University School of Graduate Studies / OhioLINK, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=case1310138396.

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18

Eriksson, Cecilia. "Affinity based proteomics research tools /." Stockholm : Skolan för bioteknologi, Kungliga Tekniska högskolan, 2009. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-11184.

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19

Mayes, Andrew Geoffrey. "Quantitative aspects of affinity adsorption." Thesis, University of Bath, 1992. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.303403.

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20

Gyepi-Garbrah, I. A. "Studies in tentacle affinity chromatography." Thesis, University of Bath, 1997. https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.362255.

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21

Senac, Caroline. "Solvants et sites de liaison hydrophobes." Thesis, Sorbonne université, 2018. http://www.theses.fr/2018SORUS021/document.

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La plupart des sites de liaisons des cibles thérapeutiques sont hydrophobes. La majorité des molécules actives sont donc hydrophobes et difficilement soluble dans l'eau. Pour remédier à ce problème lors des test in vitro, un cosolvant est ajouté dans le milieu de réaction pour augmenter la solubilité des molécules. Le plus commun est le diméthylsulfoxyde (DMSO). Cependant, peu d'études ont regardé l'effet du DMSO sur l'affinité des ligand hydrophobes pour leurs cibles, mais toutes ont montré une modification de l'affinité induite par le DMSO. Pour mieux comprendre l'effet du DMSO il est nécessaire d'étudier un grand nombres de système ligand/récepteur différents. Dans cette étude, nous avons étudié l'effet d'une concentration de 5% de DMSO sur la liaison d'un ligand hydrophobe l'acide adamantane-1-carboxylique (ADA) dans deux cavités hydrophobes la beta et la gamma cyclodextrines (CD). Les deux CD diffèrent par la largeur de leur cavité. Les mesures d'affinités ont été réalisées par mesure de vélocité ultrasonore, résonance magnétique nucléaire et par des simulations moléculaires (MS). Toutes les techniques ont montré que le DMSO n'a pas d'effet sur l'affinité de l'ADA pour la gamma-CD alors qu'il diminue fortement l'affinité de l'ADA pour la beta-CD. L'analyse des MS a montré que le DMSO entre en compétition avec l'ADA pour la petite cavité de la beta-CD alors que pour la large cavité de la gamma-CD cette compétition n'est pas présente. L'effet du DMSO semble plus important lorsque le volume du ligand est parfaitement adapté au volume de la cible
Most therapeutic targets are proteins whose binding sites are hydrophobic cavities. For this reason, the majority of drugs under development are hydrophobic molecules exhibiting low solubility in water. To tackle this issue, a few percent of cosolvent, such as dimethyl sulfoxide (DMSO), is usually employed to increase drug solubility during the drug screening process. However, the few published studies dealing with the effect of adding DMSO showed that the affinity of hydrophobic ligands is systematically underestimated. To better understand the effect of DMSO, there is a need of studying its effect on a large range of systems. In this work, we used beta and gamma-cyclodextrins (CD) as models of hydrophobic cavities to investigate the effect of the addition 5% DMSO on the affinity of 1-adamantane carboxylic acid (ADA) to these CD. The two systems differ by the size of the CD cavity. The evaluation of binding constants was performed using ultrasound velocimetry, nuclear magnetic resonance spectroscopy, and molecular simulations (MS). All techniques show that the presence of 5% DMSO does not significantly modify the affinity of ADA for gamma-CD, while the affinity is dramatically reduced for beta-CD. MS analysis show a competition between DMSO and ADA for the small cavity of beta-CD, which is not present for the large cavity of gamma-CD. The bias induced by the presence of DMSO is thus more important when the ligand volume better fits the CD cavity
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22

Garg, Nandita. "Polymer shielded dye-affinity chromatography and temperature induced phase separation two strategies to simplify protein affinity separation processes /." Lund : Dept. of Biotechnology, Lund University, 1995. http://books.google.com/books?id=Qw5rAAAAMAAJ.

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23

Singh, Jai Prakash. "Syntheses of affinity ligands and monoazaporphyrins." Thesis, University of British Columbia, 1987. http://hdl.handle.net/2429/41441.

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Heme and heme proteins are ubiquitous in nature and play many varied and important biological functions. The biological properties of the heme proteins are dependent on the interaction between the metal and the porphyrin where each is dependent on the other to carry out specific functions. This thesis describes the syntheses of affinity ligands and monoazaporphyrins. The synthesis of the ligand was based on a protoporphyr in-IX derivative substituted on either the 2- or 4-vinyl group. The two isomeric affinity ligands, 73 and 74 (2- or 4- substituted) were synthesized from protoporphyrin-IX di-tert-butyl ester 106b by conversion of the vinyl groups to monoformyl derivatives 122b (or 123b) via the intermediacy of photoprotoporphyrin-IX derivatives. The spacer chain [formula omitted] using a formyl group was extended by Knoevenagel condensation with malonic acid to give monoacrylic acid derivatives 132 (or 133). This chain was further extended by linking an aliphatic diamine (1,3-diaminopropane) to the monoacrylic acid derivatives 132 (or 133) through an acid chloride mediated amide linkage. The Fe(III) and Co(II) complexes of the affinity ligand (73 + 74) were prepared and the tertiary butyl groups were removed by treatment with trifluoroacetic acid at room temperature to give 71 and 72. [formula omitted] An improved synthesis of monoazaporphyrins (5-aza) (see p.iv) is described in this work (section 3.4). Our approach consists of constructing 1,19-dibromo-l,19-dideoxybiladiene-ac dihydrobromides which were subsequently cyclized, to give corresponding monoazaporphyrins in high yields, using dibenzo-18-crown-6 as a phase transfer agent for the transfer of an azide ion. [formula omitted]
Science, Faculty of
Chemistry, Department of
Graduate
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24

Buttery, Robert Christians. "Integrin affinity modulation and lung cancer." Thesis, University of Edinburgh, 2006. http://hdl.handle.net/1842/29025.

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Recent work has shown that the transmembrane protein CD98 is able to influence the affinity with which β1 integrins bind to extracellular ligands. The first part of this thesis presents confocal microscopy and co-immunoprecipitation experiments that confirm the physical juxtaposition of the two proteins within the cell membrane, suggesting a direct functional link between the two. It also demonstrated that cross-linking CD98 stimulates both phosphoinositide 3-kinase intracellular signalling and increased β1 integrin-dependent cellular adhesion. Because of the role of CD98 in integrin affinity modulation, the immunohistochemical expression of CD98 and its ligand, galectin-3, was studied in a variety of human ling diseases including lung cancers. The major finding of this work was a striking distinction between high expression of galectin-3 in non-small cell lung cancer and low expression in small cell lung cancer. This may hag significant implications for the differing clinical behaviours of these two groups of cancers. The final section of this thesis returns to describe experiments aimed at defining the molecular regulators of integrin affinity more clearly. A genetic screen of a cDNA library was undertaken to identify candidate genes coding for proteins able to rescue integrins from the low affinity state induced by the small signalling protein H-Ras. This identified a candidate cDNA 480, recognised to be part of a novel gene Nessie, which codes for a large protein with multiple transmembrane domains. Both 480 and Nessie appear to have the ability to rescue integrin affinity from H-Ras suppression.
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25

Lindgren, Joel. "Chemical Engineering of Small Affinity Proteins." Doctoral thesis, KTH, Proteinteknologi, 2014. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-141014.

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Small robust affinity proteins have shown great potential for use in therapy, in vivo diagnostics, and various biotechnological applications. However, the affinity proteins often need to be modified or functionalized to be successful in many of these applications. The use of chemical synthesis for the production of the proteins can allow for site-directed functionalization not achievable by recombinant routes, including incorporation of unnatural building blocks. This thesis focuses on chemical engineering of Affibody molecules and an albumin binding domain (ABD), which both are three-helix bundle proteins of 58 and 46 amino acids, respectively, possible to synthesize using solid phase peptide synthesis (SPPS). In the first project, an alternative synthetic route for Affibody molecules using a fragment condensation approach was investigated. This was achieved by using native chemical ligation (NCL) for the condensation reaction, yielding a native peptide bond at the site of ligation. The constant third helix of Affibody molecules enables a combinatorial approach for the preparation of a panel of different Affibody molecules, demonstrated by the synthesis of three different Affibody molecules using the same helix 3 (paper I). In the next two projects, an Affibody molecule targeting the amyloid-beta peptide, involved in Alzheimer’s disease, was engineered. Initially the N-terminus of the Affibody molecule was shortened resulting in a considerably higher synthetic yield and higher binding affinity to the target peptide (paper II). This improved variant of the Affibody molecule was then further engineered in the next project, where a fluorescently silent variant was developed and successfully used as a tool to lock the amyloid-beta peptide in a β-hairpin conformation during studies of copper binding using fluorescence spectroscopy (paper III). In the last two projects, synthetic variants of ABD, interesting for use as in vivo half-life extending partners to therapeutic proteins, were engineered. In the first project the possibility to covalently link a bioactive peptide, GLP-1, to the domain was investigated. This was achieved by site-specific thioether bridge-mediated cross-linking of the molecules via a polyethylene glycol (PEG)-based spacer. The conjugate showed retained high binding affinity to human serum albumin (HSA) and a biological activity comparable to a reference GLP-1 peptide (paper IV). In the last project, the possibility to increase the proteolytic stability of ABD through intramolecular cross-linking, to facilitate its use in e.g. oral drug delivery applications, was investigated. A tethered variant of ABD showed increased thermal stability and a considerably higher proteolytic stability towards pepsin, trypsin and chymotrypsin, three important proteases found in the gastrointestinal (GI) tract (paper V). Taken together, the work presented in this thesis illustrates the potential of using chemical synthesis approaches in protein engineering.

QC 20140207

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Worthington, Steve. "Affinity credit cards and relationship marketing." Thesis, Staffordshire University, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.402436.

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27

Elliott, Paul Anthony. "Integrin affinity modulation and survival signalling." Thesis, University of Edinburgh, 2008. http://hdl.handle.net/1842/4393.

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Integrins are heterodimeric transmembrane proteins that provide a bi-directional link between the cell’s internal biological mechanisms and the extracellular environment. During inside-out signalling, intracellular messages converge on the integrin cytoplasmic domain to induce a conformational change. This is transmitted to the extracellular domain where it results in an alteration in affinity for integrin ligands such as fibronectin and laminin. In this way the cell has developed the ability to modulate the critical functions of adhesion and cell movement. In outside-in signalling, the integrin performs a more complex function than simple adhesion; upon binding to ligand, the integrin extracellular domain undergoes a conformational change which is transmitted to the cytoplasmic domain. This alters the integrin’s cytoplasmic domain affinity for intracellular signalling proteins and results in the activation of intracellular second messenger pathways. In this way, the extracellular milieu is able to influence intracellular signalling including those involved in apoptosis. This thesis demonstrates data which provide original insights into bi-directional integrin signalling: Inside-out signalling: Constitutively active Notch1 increases β3-integrin affinity and abrogates Hras-mediated integrin suppression without increasing expression of β3- integrin. Dominant-Negative Rras blocks Notch-mediated integrin activation and Notch1-mediated reversal of Hras and Raf-mediated integrin suppression and this is independent of erk phosphorylation. Notch1 induces Rras activation. Functional adhesion assays confirm that Notch1IC increases K562 adhesion in a β1-integrin dependent manner and this is abrogated by Dominant-Negative Rras. This data supports a mechanism in which Notch1 increases integrin affinity via activation of Rras. Outside-in signalling: Evidence is presented demonstrating that extracellular matrix proteins, laminin and fibronectin, activate β1-integrins to protect SCLC cells against the apoptotic effects of etoposide and ionizing radiation via PI3Kinase activation. This occurs in two ways: 1) PI3Kinase-dependent β1-integrin signalling resulting in phosphorylation of Bad and reduced caspase-9 cleavage and 2) a β1-integrinmediated over-riding of etoposide and radiotherapy-induced cell cycle S phase delay and G2/M arrest. β1-integrin-mediated outside-in survival signalling was investigated further in the in vivo setting; MatrigelTM, a basement membrane product rich in extracellular matrix proteins, promoted SCLC xenograft survival and growth in a β1-integrin and tyrosine kinase-dependent manner. This data provides novel insights into the critical functions that integrins play in adhesion and survival signalling.
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Forde, G. M. "Plasmid DNA purification by affinity methods." Thesis, University of Cambridge, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.599116.

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Two affinity mechanisms were investigated for their suitability to pDNA purification: (1) GSH/GST-ZnF/pTS: a dual affinity fusion protein, comprised of Glutathione-S-Transferase (GST) and a zinc finger transcription factor (ZnF), was utilised to purify target pDNA. The fusion protein was firstly adsorbed to an immobilised glutathione (GSH) ligand via the GST segment followed by specific adsorption of a pUC19 based plasmid (pTS) to the exposed zinc finger; and (2) lac I peptide/pLS3: a lac repressor (lacI) peptide that displays affinity for lac operator (lacO) sequences was used to purify target pDNA. A pUC19 based plasmid containing lacO sequences (pLS3) was adsorbed to the immobilised lacI peptide. The lac I peptide/pLS3 mechanism showed characteristics that make it more applicable to the commercial purification of pDNA. These included: a simpler adsorption mechanism (i.e. no intermediate affinity molecule such as GST-ZnF was required), high ligand utilization, the peptide ligand can be directly immobilised to an adsorbent matrix, and high purity pDNA can be eluted directly into a formulation buffer suitable for pDNA storage (i.e. 10 mM Tris Base, 1 mM EDTA, pH 8) so that no further processing is required. The major disadvantages of the system are low elution yields (34 %) and the cost of producing the affinity ligand. These draw backs may be reduced by designing a peptide ligand that does not bind pDNA as strongly and is less expensive to produce (i.e. shorter or synthetic). Through process innovation (utilisation of an affinity binding mechanism in scalable unit operations), the potential for affinity chromatography as a process for pDNA purification was realised.
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Smith, Jane H. "Personality types and affinity for computers." Thesis, Monterey, California. Naval Postgraduate School, 1991. http://hdl.handle.net/10945/26691.

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Trevitt, Clare Rosalind. "Designing high affinity ligands for calmodulin." Thesis, University of Sheffield, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.299601.

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Brown, Robert Gareth Sumser. "The affinity labelling of gibberellin hydroxylases." Thesis, University of Bristol, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.295169.

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Foaden, Shaun Patrick. "Continuous affinity partitioning for protein purification." Thesis, University of Cambridge, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.335093.

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Pattison, R. "Characterisation of next generation affinity reagents." Thesis, University of Liverpool, 2017. http://livrepository.liverpool.ac.uk/3021754/.

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Brown, Alec J. "Ipsative Score Distortion on Affinity 2.0." Diss., CLICK HERE for online access, 2005. http://contentdm.lib.byu.edu/ETD/image/etd1119.pdf.

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Noland, Timothy L. "Affinity-seeking and superior-subordinate communication /." free to MU campus, to others for purchase, 1998. http://wwwlib.umi.com/cr/mo/fullcit?p9901266.

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Blomberg, Lennart. "Synthesis, coupling and use of oligosaccharides in affinity chromatography." Lund : Organic Chemistry 2, Lund Institute of Technology, Lund University, 1994. http://books.google.com/books?id=-A1rAAAAMAAJ.

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Marillet, Simon. "Modélisation de la réponse des anticorps : de la structure des complexes immunoglobuline - antigène à la complexité clonale des répertoires de chaines lourdes d'immunoglobulines." Thesis, Université Côte d'Azur (ComUE), 2016. http://www.theses.fr/2016AZUR4120/document.

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Cette thèse étudie trois sujets relevant de la biologie structurale, de lagénétique et de l'immunologie.Premièrement, nous développons de nouveaux prédicteurs de l'affinité deliaison de complexes protéiques, produisant des résultats de niveau ``état del'art''. Nous calculons d'abord 12 variables modélisant diverses propriétésstructurales des complexes. Nous générons et évaluons des estimateursutilisant des sous ensembles de ces variables, de façon à identifier les plusperformants. Le logiciel associé est distribué dans la Structural BioinformaticsLibrary.Deuxièmement, nous proposons de nouvelles analyses de complexes Ig-Ag.D'une part nous concevons un classificateur distinguant les types de ligand desIg. D'autre part, nous montrons que le modèle précédent prédit fidèlementl'affinité de complexes Ig-Ag. Enfin, nous quantifions la contribution des CDR3de la chaine lourde à l'affinité de liaison, et montrons qu'il contribuesignificativement plus que les autres CDR.Enfin, nous nous intéressons à la modélisation de la diversité des répertoiresde chaîne lourde des Igs, à partir de données de séquençage de CDR3, dans unmodèle de vaccin chez le poisson. Nous analysons les répertoires dans troisconditions: naifs, vaccinés et vaccinés + infectés. Nous comparons lesrépertoires de deux individus en utilisant la « earth-mover distance », laquelleexploite la correspondance entre clonotypes de deux répertoires, révélant ainsides informations inaccessibles aux méthodes basées sur les indices dediversité.Dépôt de thèseDonnées complémentairesPour caractériser la notion de réponse immunitaire publique / privée, nousquantifions le chevauchement des clonotypes exprimés entre individus de lamême ou de différentes conditions
This thesis investigates three topics at the cross-roads of structural biology,genetics and immunology.First, we develop a pipeline to design and select binding affinity predictors forprotein complexes, yielding state-of-the art results. The first step is the designand computation of 12 different variables accounting for geometric andphysico-chemical properties of the complexes. The second step is thegeneration and evaluation of models using subsets of these variables, followedby the selection of the best performing ones. The corresponding software isdistributed within the Structural Bioinformatics Library.Second, we provide an analysis of the interface properties of Ig-Ag complexes.In particular, we design a classifier using two descriptors, which is able todistinguish ligand types. We also apply the previous binding affinity predictionmodel to Ig-Ag complexes and obtain accurate predictions. We then develop aquantitative model for the contribution of VH CDR3 to the binding affinity andinteraction specificity, and show that it contributes significantly more thanother CDRs.Third, we model the diversity of VH CDR3 repertoires from Ig RNA sequencingdata in a fish vaccination model. We analyze repertoires from three conditions:naive, vaccinated and vaccinated + infected fish. Comparison of the repertoiresof two individuals uses the earth-mover distance (EMD). By exploiting amapping between the clonotypes of the repertoires, we show that EMD revealsinformation beyond classical methods based on diversity indexes. Tocharacterize the notion of public / private immune response, we quantify theoverlap of clonotypes between individuals of the same or different conditions
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38

Göbel, Christian. "Strategisches Affinity-group-Netzwerkmanagement Risikoreduzierung mittels koevolutionärer Anpassung an und Gestaltung von szenebasierten Netzwerkstrukturen aus fokaler Unternehmenssicht." Hamburg Kovač, 2009. http://d-nb.info/995566704/04.

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39

Sundberg, Mårten. "Protein microarrays for validation of affinity binders." Licentiate thesis, KTH, Proteomik, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-48256.

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Is specificity an important issue regarding affinity reagents? What about the validation of affinity reagents today, is it good enough? This depends on the application and the producer of the reagent. Validation should be the most important marketing argument that can be found.Today there is a continuous growth of both the number of affinity reagents that are produced and the different types of affinity reagents that are developed. In proteomics they become more and more important in exploring the human proteome. Therefore, validated affinity reagents should be on top of every proteomic researcher’s list. How should this be accomplished?Better international agreements on how affinity reagents should be tested to be regarded as functional reagents are needed. One of the most important issues is the specificity of the affinity reagent. An international standard for which specific validation that is needed for different kinds of applications would be very useful.In this thesis, it is shown that the protein microarray platform that was established within the HPA project at KTH is a very good tool to determine the specificity of different affinity binders.In the first study, the production of mono-specific antibodies for tissue profiling in the Human Protein Atlas (HPA) project is presented. The section describing the use of protein microarrays for validation of the antibodies is relevant for this thesis. The implementation of protein microarrays in the HPA workflow was an important addition, because a deeper insight of the specificity of all the antibodies produced were now available.In a second study, bead based arrays were compared to planar protein microarrays used in the HPA project. In this study, 100 different bead identities were coupled with 100 different antigens and mixed together to generate an array. The correlation between the two types of assays was very high and the conclusion was that the methods can be used as backup to each other.A third study was a part of an international initiative to produce renewable affinity binders against proteins containing SH2 domain. Here, the HPA protein microarrays were modified to analyze different types of reagents produced at six laboratories around the world. Monoclonal antibodies, single chain fragment and fibronectin scaffolds were tested as well as mono-specific antibodies. It was shown to be possible to adapt protein microarrays used in the HPA project to validate other kinds of affinity reagents.
QC 20111117
Development and applications of protein microarrays
The Swedish Human Proteome Resource (HPR) program
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40

Lad, Yatishkumar. "Integrin affinity modulation by Ras signalling molecules." Thesis, University of Edinburgh, 2002. http://hdl.handle.net/1842/24800.

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In this thesis I have sought to understand the mechanism by which H-Ras and its effectors modulate integrin affinity. H-Ras is a member of the Ras superfamily of small GTP binding proteins. Expression of the constitutively active variant of H-Ras (Ras G12V) within an integrin affinity reporter system (αβ-py cells) reduced integrin affinity (suppressed integrin). Ras effector mutants revealed that integrin suppression is mediated by Raf-dependent and Raf-independent signalling pathways. Raf-independent signalling pathways activated by Ral-GEFs and PI3-kinase were not recognisable for integrin suppression. An active variant of R-Ras (R-Ras G38V) reversed integrin suppression by both Raf-dependent and - independent pathways, indicating that these pathways may converge at a point proximal to the integrin. Raf initiates a protein signalling cascade leading to ERK activation that is responsible for many of the Ras/Raf-dependent biological functions. However, Raf-dependent integrin suppression was insensitive to MEK inhibition with the PD098059 compound. A novel Raf mutant (T481A) that fails to bind to MEK was also capable of mediating integrin suppression in the absence of ERK activation. Surprisingly, Raf-BxB T481A-mediated integrin suppression was sensitive to expression of MKP-1. Taken together it is proposed that Raf may mediate integrin suppression via a MEK-independent pathway that may utilise a member of the MAP kinase superfamily. In conclusion, integrin suppression by Ras is mediated by both Raf-independent and dependent pathways. Signalling by Raf may utilise components other than those present in the classical Ras to ERK protein cascade.
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41

Tarasova, Anna Optometry UNSW. "Fabrication and characterisation of affinity-bound liposomes." Awarded by:University of New South Wales. Optometry, 2007. http://handle.unsw.edu.au/1959.4/29114.

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In considering the concept of surface-immobilised liposomes as a drug release system, two factors need to addressed, the interfacial surface density of the liposomes for maximum drug loading and the stability of these liposomes to allow for controlled drug release. This thesis investigates a multilayer system for the affinity immobilisation of liposomes and their stability to various applied stresses. In the work presented here an allylamine monomer was used to create plasma coatings that were stable, thin and amine-rich. The aging studies using AFM showed these films to rapidly oxidise on exposure to water. The freshly deposited films were used for further surface modifications, by the covalent grafting of PEG layers of different interfacial densities under the conditions of varying polymer solvation. The AFM was used to measure the interaction forces between the grafted PEG layers and modified silica interfaces. It was found that the polydispersity of the PEG species resulted in bridging interactions of ???brush???-like PEG layers with the silica surface. These interactions were screened minimised by increasing the ionic strength of the solution. Although the densely grafted PEG layers were found to be highly protein-resistant by the XPS and QCM-D some minor protein-polymer adhesions were observed by the AFM. The densely anchored biotinylated PEG chains served as an optimum affinity platform for affinity-docking of NeutrAvidinTM molecules, which assembled in a rigid, 2-D layer as confirmed by the QCM-D. The submonolayer surface density of NeutrAvidin, as determined by Europium-labelling, was attributed to steric hindrance of the immobilised molecules. The final protein layer enabled specific binding of biotin-PEG-liposomes as a highly dissipative, dense and stable layer verified by tapping mode AFM and QCM-D. We found that these liposomes were also stable under a range of stresses induced by the shearing effects of water, silica probe and HSA layer at increased loads and velocities. The frictional response of the liposome layer also demonstrated the viscoelasticity and stability of these surface immobilised liposomes. Finally, the minimal adhesive interaction forces, as measured by the AFM, demonstrated the repellency of these liposomes to commonly found proteins, such as HSA.
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42

Henderson, J. S. "Combined microfiltration and membrane-based affinity separation." Thesis, University of Oxford, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.325959.

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43

Stephenson, Heather. "Screening and Diagnostic Validity of Affinity 2.5." Thesis, Brigham Young University, 2015. http://pqdtopen.proquest.com/#viewpdf?dispub=3680970.

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Affinity 2.5 is a computer-based instrument designed to assess sexual interest using viewing-time measures. Viewing-time measures of sexual interest have been developed to identify individuals with deviant sexual interest. The purpose of this study is to examine the validity of Affinity 2.5 in screening and diagnosing individuals with sexually deviant interests. This study used viewing time profiles of known sexual offenders compared to norm-referenced profiles of an exclusively heterosexual, non-pedophilic college population. Participants were 155 males and 3 females who had sexually offended against children and 63 male and 84 female non-offender college students. Results show that 43.7% of offenders were correctly identified as having significantly deviant sexual interest, compared to the reference group. Further 12.0% of offenders showed statistical significant interest in at least one category of individuals from a protected population and offended against that same category. The results of this study do not provide support for the utility of the Affinity 2.5 as a screening or diagnostic tool.

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44

Bradshaw, Anthony Paul. "Modification of polysaccharides in affinity precipitation studies." Thesis, Heriot-Watt University, 1990. http://hdl.handle.net/10399/901.

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45

Gallacher, Stuart. "Studies on the affinity precipitation of proteins." Thesis, Heriot-Watt University, 1994. http://hdl.handle.net/10399/1416.

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Hawkins, Robert Edward. "New methods for selecting high affinity antibodies." Thesis, Open University, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.240367.

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47

McCreath, Graham Edward. "Development and applications of perfluorocarbon affinity emulsions." Thesis, University of Cambridge, 1993. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.307052.

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48

Miller, Katherine Florence. "Fabrication and processing of affinity tagged nanoparticles." Thesis, University of Cambridge, 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.612279.

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Bruger, Annika Målin. "TCR signalling in response to affinity stimulation." Thesis, University of Oxford, 2013. http://ora.ox.ac.uk/objects/uuid:5f9c1001-6c43-472f-a495-c9573b54a84a.

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T cells are an essential part of the adaptive immune system and protect the body from intracellular infections. The specificity with which αßTCR-bearing T cells recognize cognate antigen presented on MHC molecules is paramount to maintaining the balance between mounting effector functions against pathogens and establishing peripheral tolerance to self. The mechanism by which T cells translate qualitative differences in TCR:pMHC binding to sensitive proximal signalling events which ultimately result in specific Tcell effector responses to infected cells but not to self is mostly unknown. To address how T cell signalling responds to qualitative differences in TCR triggering by pMHC, I established a system of stimulating T cells bearing the 1G4 TCR specifically in vitro with a panel of four NY-ESO-1156-165 peptide variant MHC tetramers. Single amino acid substitutions to the NY-ESO-1156-165 peptide conferred a maximum 35-fold difference in the monomeric affinity for the 1G4 TCR. The system allows the highly controlled investigation of very rapid TCR proximal signalling events simultaneously and quantitatively using flow cytometry. Stimulations with pMHC tetramers showed rapid sensitive sequential signalling responses which were able to confer ligand discrimination. Very early signalling events such as CD3ζ phosphorylation showed analogue responses to the different affinity pMHC tetramers. Later signalling events including phospho-ERK showed a distinct on/off switch-like response. The amplitude of the very early analogue signalling responses determined the extent of later digital ERK signals. This indicates that a certain analogue signalling threshold must be passed to result in T cell activation. The thymocyte protein Themis has been shown proximal TCR signalling to modulate thymocyte selection thresholds. Its deletion results in profound defects in positive thymocyte selection. Themis locates to the LAT signalosome of the TCR signalling cascade via Grb2, yet its molecular function is unknown. Employing the system I established, I demonstrate that Themis-k/d cells show increased levels of CD3z-chain phosphorylation, phospho-ERK signalling and signal-induced apoptosis which was independent of the ERK signal. This shows that Themis is a global attenuator of proximal TCR signalling. We are currently investigating possible associations of Themis to proteins phosphastases such as SHP-1 which could attenuate TCR proximal protein tyrosine signalling events.
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50

Barroso, Telma Godinho. "Preparation of affinity membranes using alternative solvents." Master's thesis, Faculdade de Ciências e Tecnologia, 2008. http://hdl.handle.net/10362/7875.

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