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Dissertations / Theses on the topic 'Proteine antigelo'

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Published: 9 March 2023
Last updated: 10 March 2023

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1

MANGIAGALLI, MARCO. "Structural and functional analyses of an ice-binding protein from an Antarctic bacterium." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2019. http://hdl.handle.net/10281/241269.

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Una proteina in grado di legare i cristalli di ghiaccio è definita proteina legante il ghiaccio o IBP acronimo dall’inglese ice-binding protein. Le IBP grazie alla loro capacità di abbassare il punto di congelamento dell’acqua, aumentando il gap di isteresi termica (TH). Questo intervallo è definito come la differenza tra il punto di fusione e di congelamento dell’acqua. La seconda attività delle IBP è l’inibizione della ricristallizzazione del ghiaccio (ice recrystallization inhibition, IRI). Infatti, queste proteine stabilizzano i piccoli cristalli di ghiaccio impedendo la formazione di cristalli di ghiaccio di grosse dimensioni che sono dannosi per le cellule. Le IBP sono state identificate in numerosi organismi tra cui pesci, insetti, batteri, alghe e lieviti. Queste proteine rappresentano un esempio di evoluzione convergente, infatti tutte le IBP condividono lo stesso meccanismo di legame con il ghiaccio nonostante una sorprendente diversità strutturale e funzionale. Questo lavoro di tesi è focalizzato sulla caratterizzazione funzionale e strutturale di EfcIBP, una IBP batterica identificata da analisi di metagenomica effettuate sul ciliato Antartico Euplotes focardii e sul consorzio batterico ad esso associato. La struttura 3D di EfcIBP è stata risolta mediante cristallografia ai raggi X e consiste in un β-solenoide con un α-elica parallela all’asse principale della proteina. L’analisi strutturale ha permesso di identificare tre diverse facce del solenoide denominate A, B e C. Simulazioni di docking suggeriscono che EfcIBP è in grado di legare i cristalli di ghiaccio tramite le facce B e C del solenoide. Questa ipotesi è stata verificata attraverso la progettazione razionale di 6 varianti che sono state prodotte e saggiate per la loro attività. In generale, questi risultati indicano che EfcIBP è in grado di legare i cristalli di ghiaccio attraverso le facce B e C del solenoide. Questa peculiarità strutturale si riflette in un’insolita combinazione di attività di IRI e TH. Infatti, EfcIBP presenta una notevole attività di IRI in un intervallo di concentrazione nanomolare e una attività di isteresi termica di 0.53°C alla concentrazione di 50 μM che la rende una IBP moderata. All’interno del gap di TH, i cristalli di ghiaccio presentano una forma esagonale, mentre a temperature al di sotto della temperatura di congelamento presentano una forma a “Saturno". La proteina chimerica formata dalla “green fluorescent protein” e da EfcIBP è stata utilizzata per determinare a quali piani del cristallo di ghiaccio la proteina è in grado di legarsi e con quale cinetica. I dati sperimentali suggeriscono che le peculiarità funzionali di EfcIBP sono dovute alla sua capacità di legare velocemente i piani basali e piramidali del cristallo di ghiaccio. Questi dati, insieme alla presenza di una sequenza segnale per la secrezione, suggeriscono che EfcIBP è secreta e svolge la funzione di mantenere liquido l’ambiente circostante aumentando lo spazio vitale. In conclusione, EfcIBP è un nuovo tipo di IBP con proprietà insolite di legame al ghiaccio e di attività di IRI. Questo studio ha contribuito ad identificare una nuova classe di IBP moderate che potrebbero essere sfruttate come crioprotettori in diversi settori come la criobiologia e quello alimentare.
Ice-binding proteins (IBPs) are characterized by the ability to control the growth of ice crystals. IBPs are active in increasing thermal hysteresis (TH) gap as they decrease the freezing point of water. On the other hand, IBPs can inhibit ice recrystallization (IRI) and stabilize small ice crystals at the expense of the harmful, large ones. IBPs have been identified in several organisms including higher Eukaryotes and microorganisms such as bacteria, yeasts and algae. Although IBPs share the ability to bind ice crystals, proteins from different sources present different 3D structures, from α-helix to β-solenoid proteins. This thesis is focused on the structural and functional characterization of EfcIBP, a bacterial IBP identified by metagenomic analysis of the Antarctic ciliate Euplotes focardii and the associated consortium of non-cultivable bacteria. The 3D structure of EfcIBP, solved by X-ray crystallography, consists in a β-solenoid with an α-helix aligned along the axis of the β-helix. It is possible to distinguish three different faces: A, B and C. Docking simulations suggest that B and C faces are involved in ice binding. This hypothesis was tested by the rational design of six variants that were produced and assayed for their activity. Overall, these experiments indicate that both solenoid faces contribute to the activity of EfcIBP. EfcIBP displays remarkable IRI activity at nanomolar concentration and a TH activity of 0.53°C at the concentration of 50 μM. The atypical combination between these two activities could stem from the ability of this protein to bind ice crystals through two faces of the solenoid. In the presence of EfcIBP, ice crystals show a hexagonal trapezohedron shape within the TH gap, and a unique “Saturn-shape” below the freezing point. A chimeric protein consisting of the fusion between EfcIBP and the green fluorescent protein was used to deeper investigate on this aspects by analyses of fluorescence ice plane affinity and binding kinetics. Overall, experimental data suggest that the EfcIBP unique pattern of ice growth and burst are due to its high rate of binding at the basal and the pyramidal near-basal planes of ice crystals. These data, together with the signal sequence for the secretion, suggest that EfcIBP is secreted in local environment where it becomes active in increasing the habitable space. In conclusion, EfcIBP is a new type of IBP with unusual properties of ice shaping and IRI activity. This study opens new scenarios in the field of IBPs by contributing to identify a new class of moderate IBPs potentially exploitable as cryoprotectants in several fields, such as cryobiology and food science.
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2

Varelias, Antiopi. "Studies of CD44 variant isoform expression and function on activated human peripheral blood mononuclear cells and in renal transplantation." Title page, summary and contents only, 2001. http://web4.library.adelaide.edu.au/theses/09PH/09phv293.pdf.

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3

Winchester, Christopher Charles. "The roles of Hsp70 proteins in antigen processing and presentation." Thesis, University of Oxford, 1997. http://ora.ox.ac.uk/objects/uuid:567dff45-08ce-43b4-b011-d08afea42f76.

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The ability of members of the hsp70 family to bind to peptides in vivo and in vitro suggests that they may be involved in the processing of antigens for binding to Major Histocompatibility (MHC) class I and/or class n molecules. The aims of this thesis have been to provide evidence for the involvement of hsp70s in antigen processing and to characterise the binding of peptides by hspTOs by structural and functional studies. Firstly, the peptide-binding domains of two hsp70s, hsp70hom and PBP74, were expressed in isolation from the rest of the molecule for structure determination. Both of these hsp70s were implicated in antigen processing: hsp70hom in the class I pathway, due to its cytoplasmic localisation and constitutive expression, and the presence of its gene in the MHC; and PBP74 in the class n pathway because published work indicated that it was localised to endosomes and that antibodies against it inhibited antigen processing. The expression and purification of both peptide-binding domains was very successful, and one dimensional NMR experiments indicated that they were folded. However, it was not possible to determine their structures by NMR spectroscopy or X-ray crystallography because they aggregated in solution at high concentrations. Instead, the structure of the C-terminal region of hsp70hom, which includes its peptidebinding domain, was modelled based on the known structure of the equivalent portion of dnaK, the hsp70 of E.coli. The structure of hsp70hom is predicted to be very similar to that of dnaK, and modelling studies suggest that it is likely to bind peptides in a closely related fashion. The modelling of complexes between hsp70hom and two peptides suggest that the peptide-binding groove is very versatile, accounting for the broad peptide-binding specificity of hsp70s. The interactions of hsp70hom and PB74 with peptides were investigated using plate binding assays and isothermattitration calorimetry. A biotinylated peptide bound to the peptide-binding domain of hsp70hom, immobilised in plastic wells, with a Kd of <25 μM, which is within the range of Kds reported for other hsp70-peptide complexes (0.1-100 μM). In solution, isothermal titration calorimetry showed that the binding of peptides to the peptide-binding domains of hsp70hom and PBP74 was likely to be entropically rather than enthalpically driven, and, therefore, the interactions involved are likely to be predominantly hydrophobic. Secondly, PBP74, an hsp70 thought to be involved in the class II antigen processing pathway in endosomes, was localised by immunofluorescence microscopy. It was shown to be a mitochondrial protein, and is, therefore, unlikely to be involved in antigen processing. The presence of other members of the hsp70 family in lysosomes purified from a B cell line by Percoll density gradient centrifugation was investigated using antibodies that reacted with many Afferent members of the hsp70 family. No hsp70s were detected in these late endocytic compartments, even after heat shock or serum starvation. However, the presence of an hsp70 in endosomes, or of a member of this family not detected by the antibodies used, in lysosomes, cannot be ruled out. A third approach investigated the induction of the three hsp70 genes found in the MHC by four cytokines. The hsp70-l and hsp70-2 genes are induced at the mRNA level by IFN-γ and IL- 1, while TNF induces hsp70-2 alone. This data supports a role for the heat-inducible hsp70 in MHC class I antigen processing, as it appears to be coregulated with known members of this antigen processing pathway. The expression of hsp70hom was unaffected by any of the four cytokines examined. In addition, the mitochondrial hsp70 (which is not encoded in the MHC) appears to be induced by IFN-γ at the protein level. The research presented in this thesis provides a greater understanding of the peptide-binding properties of two hsp70s. Further work is necessary to show conclusively whether any of the hsp70s is involved in antigen processing.
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4

Malhotra, Shikha. "B-cell-antigen receptor endocytosis uses a distinct signaling pathway, involving LAB, Vav, dynamin and Grb2." Oklahoma City : [s.n.], 2009.

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5

MENTO, ALFREDO. "Unconventional purification and labelling strategies of bioreagents for immunodiagnostic assays." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2021. http://hdl.handle.net/10281/309986.

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Gli antigeni e gli anticorpi sono reagenti chiave per lo sviluppo di test immunodiagnostici accurati, riproducibili e sensibili, ampiamente utilizzati per la rilevazione di malattie infettive (HIV, HBV, HCV, ecc.) e per la determinazione di marcatori biologici (vitamine, ormoni, ecc.). Questi bioreagenti devono essere prodotti con un alto grado di purezza, in formulazioni stabili e con sufficiente riproducibilità nel tempo (consistenza da lotto a lotto). Un aspetto spesso trascurato nella produzione di questi reagenti è il loro costo, che deve essere sufficientemente basso da non avere un impatto significativo sul prezzo finale dei saggi immunochimici. Le fasi di purificazione e di marcatura di questi bioreagenti incidono principalmente sul loro costo complessivo poiché vengono utilizzati reagenti e strumentazioni costosi e protocolli complessi che richiedono tempo. Per tutti questi motivi è importante ricercare nuove strategie di purificazione che permettano lo sviluppo di processi più semplici, con minor quantità di reagenti, in tempi più brevi, in modo da ridurre i costi. Pertanto, è necessario sviluppare purificazioni innovative e protocolli di marcatura sito-specifici. Nella prima parte di questo progetto abbiamo sfruttato il sistema ELP-intein. Questo metodo si basa sulla combinazione di due tools tecnologici, Elastin-like-polypeptides (ELP) (tool fisico-chimico) e l'attività dell’inteina MxeGyrA (tool biochimico), appartenente alla famiglia delle cis inteine. Ci siamo concentrati sulla purificazione e la marcatura dell'antigene C33 appartenente al virus dell'epatite C (HCV). L'antigene C33, attualmente utilizzato nel test Diasorin LIAISON® XL Murex HCV per la rilevazione di anticorpi umani contro il virus dell'epatite C, è stato purificato utilizzando un metodo non convenzionale senza passaggi cromatografici. Inoltre, abbiamo realizzato una biotinilazione sito-specifica dell'antigene C33 al suo C-terminale durante la purificazione, sfruttando l'attività biologica inteina di MxeGyrA. Sono stati sviluppati due diversi protocolli; entrambi hanno portato all'ottenimento di un antigene C33 biotinilato con un’elevata purezza e immunoreattività paragonabile a quella dell’antigene attualmente utilizzato nel saggio Diasorin HCV. Alla luce di questi buoni risultati, nella seconda parte del progetto, abbiamo studiato la possibilità di applicare la tecnologia del Protein Trans Splicing (PTS) per eseguire la marcatura sito-specifica di bioreagenti. La tecnologia PTS sfrutta l'attività delle split inteine. In particolare, nei nostri esperimenti abbiamo utilizzato la Cfa split-intein che deriva da un processo di mutagenesi della split-intein naturale Npu che ne ha notevolmente migliorato la cinetica di PTS, stabilità termica e tolleranza agli agenti caotropici. Questa nuova tecnica ci ha permesso di impostare un protocollo di marcatura sito-specifica per la produzione di bioreagenti biotinilati. Sono state utilizzate due proteine modello: lo stesso antigene C33 e una IgG umana ricombinante. Anche l'uso della tecnica PTS ha permesso di ottenere per entrambe le due proteine un'elevata purezza e prestazioni comparabili nei relativi saggi immunodiagnostici. In sintesi, il sistema ELP-inteina ha permesso di purificare l'antigene C33 senza passaggi cromatografici e di marcare in modo sito-specifico la stessa proteina al C-terminale. Inoltre, attraverso l'utilizzo del sistema Cfa split-intein abbiamo ottenuto la biotinilazione sito-specifica dell'antigene C33 e dell'IgG ricombinante. Un aspetto molto rilevante è che tutte queste proteine sono funzionali nella piattaforma LIAISON. In futuro, questi protocolli potrebbero essere utilizzati per la purificazione e / o la marcatura sito-specifica di nuovi bioreagenti utili per lo sviluppo di saggi immunodiagnostici.
Antigens and antibodies are key reagents for the development of accurate, reproducible and sensible immunodiagnostic assays, which are widely used for the detection of infectious diseases (HIV,HBV, HCV, etc.) and the determination of biological markers (vitamins, hormones, etc.). These bioreagents need to be produced at a high purity degree, in stable formulations and with sufficient reproducibility over time (lot to lot consistency). An aspect often overlooked in the production of these reagents is their cost, which must be low enough to not have a significant impact on the final price of the immunochemical assays. The purification and labeling steps of these bioreagents mainly affect the overall cost of them since costly reagents and instrumentations and complex and time-consuming protocols are used.For all these reasons it is important to seek new purification strategies that allow the development of simpler processes, with less use of reagents and shorter protocol times, and at the end minor costs. Therefore, it is necessary to develop innovative purifications and site-specific labelling protocols. In the first part of this project we exploited the ELP-intein system. This method is based on the combination of two technological tools, the Elastin-like-polypeptides (ELPs) (a physico-chemical tool) and the MxeGyrA intein activity (a biochemical tool), belonging to the cis intein family. We focused on the purification and labelling of the C33 antigen from Hepatitis C Virus (HCV). The C33 antigen, which is currently used in the Diasorin LIAISON® XL Murex HCV assay for the detection of human antibodies against the Hepatitis C Virus, was purified using a not conventional purification method without chromatographic steps. Moreover, we realized a site-specific biotinylation of C33 antigen at its C-terminus during the purification exploiting the MxeGyrA intein biological activity. Two different protocols were developed; both of them brought to the obtainment of a biotinylated C33 antigen with high purity and a comparable immunoreactivity with the one currently used in the Diasorin LIAISON® XL Murex HCV assay. In light of these good results, in the second part of the project, we investigated the possibility to apply the Protein Trans Splicing (PTS) technology to perform site-specific labelling of bioreagents. PTS technology exploits the split intein activity. In particular, in our experiments we used the Cfa split-intein which derives from a mutagenesis process of the natural Npu split-intein that significantly improved its kinetic of PTS, thermal stability and tolerance at the chaotropic agents. This new technique allowed us to set up a site-specific labelling protocol for the production of biotinylated bioreagents. Two model protein were used: the same C33 antigen and a recombinant human IgG. Also the use of PTS technique permitted to obtain for both of two proteins a high purity and a comparable performance in the immunoassays. In summary, the ELP-intein system allowed to purify the C33 antigen without chromatographic steps and then to site-specific label the same protein at the C-terminus. Moreover, through the use of the Cfa split-intein system we obtained the site-specific biotinylation of the C33 antigen and the recombinant IgG. A very relevant aspects is that all these proteins are functional in the LIAISON platform. In the future, these protocols could be used for the purification and-or the site-specific labelling of new bioreagents useful for the development of immunodiagnostic assays.
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6

Lot, Perrine. "Les protéines antigel." Paris 5, 1988. http://www.theses.fr/1988PA05P219.

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7

Schumacher, Dominik. "Site-specific functionalization of antigen binding proteins for cellular delivery, imaging and target modulation." Doctoral thesis, Humboldt-Universität zu Berlin, 2017. http://dx.doi.org/10.18452/18547.

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Antikörper und Antigen-bindende Proteine, die an Fluorophore, Tracer und Wirkstoffe konjugiert sind, sind einzigartige Moleküle, welche die Entwicklung wertvoller diagnostischer und therapeutischer Werkzeuge ermöglichen. Allerdings ist der Konjugationsschritt sehr anspruchsvoll und trotz intensiver Forschung noch immer ein bedeutender Engpass. Zusätzlich sind Antigen-bindende Proteine oftmals nicht dazu in der Lage, die Zellmembran zu durchdringen und im Zellinneren nicht funktionsfähig. Daher ist ihre Verwendung auf extrazelluläre Targets beschränkt, was eine bedeutende Anzahl wichtiger Antigene vernachlässigt. Beide Limitierungen bilden Kernaspekte dieser Arbeit. Mit Tub-tag labeling wurde ein neuartiges und vielseitiges Verfahren für die ortsspezifische Funktionalisierung von Biomolekülen und Antigen-bindenden Proteinen entwickelt, und so die Palette der Proteinfunktionalisierungen bedeutend erweitert. Tub-tag wurde erfolgreich für die ortsspezifische Funktionalisierung verschiedener Proteine und Antigen-bindender Nanobodies angewendet, die für konfokale Mikroskopie, Proteinanreicherung und hochauflösende Mikroskopie eingesetzt wurden. In einem weiteren Projekt wurden zellpermeable Antigen-bindende Nanobodies hergestellt und somit das schon lange Zeit bestehende Ziel, intrazelluläre Targets durch in vitro funktionalisierte Antigen-bindende Proteine zu visualisieren und manipulieren, erreicht. Hierzu wurden zwei verschiedene Nanobodies an ihrem C-Terminus cyclischen zellpenetrierenden Peptiden unter Verwendung von Expressed Protein Ligation funktionalisiert. Diese Peptide ermöglichten die Endozytose-unabhängige Aufnahme der Nanobodies mit sofortiger Bioverfügbarkeit. Mit Tub-tag labeling und der Synthese von zellpermeablen Nanobodies konnten wichtige Bottlenecks im Bereich der Proteinfunktionalisierung und Antikörperforschung adressiert werden und neue Tools für die biochemische und zellbiologische Forschung entwickelt werden.
Antibodies and antigen binding proteins conjugated to fluorophores, tracers and drugs are powerful molecules that enabled the development of valuable diagnostic and therapeutic tools. However, the conjugation itself is highly challenging and despite intense research efforts remains a severe bottleneck. In addition to that, antibodies and antigen binding proteins are often not functional within cellular environments and unable to penetrate the cellular membrane. Therefore, their use is limited to extracellular targets leaving out a vast number of important antigens. Both limitations are core aspects of the presented thesis. With Tub-tag labeling, a novel and versatile method for the site-specific functionalization of biomolecules and antigen binding proteins was developed expanding the toolbox of protein functionalization. The method is based on the microtubule enzyme tubulin tyrosine ligase. Tub-tag labeling was successfully applied for the site-specific functionalization of different proteins including antigen binding nanobodies which enabled confocal microscopy, protein enrichment and super-resolution microscopy. In addition to that, cell permeable antigen binding nanobodies have been generated constituting a long thought goal of tracking and manipulating intracellular targets by in vitro functionalized antigen binding proteins. To achieve this goal, two different nanobodies were functionalized at their C-terminus with linear and cyclic cell-penetrating peptides using expressed protein ligation. These peptides triggered the endocytosis independent uptake of the nanobodies with immediate bioavailability. Taken together, Tub-tag labeling and the generation of cell-permeable antigen binding nanobodies strongly add to the functionalization of antibodies and their use in biochemistry, cell biology and beyond.
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8

Heinrich, Garrett. "A role for CEACAM proteins in energy balance and peripheral insulin action." Toledo, Ohio : University of Toledo, 2010. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=mco1272976279.

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Dissertation (Ph.D.)--University of Toledo, 2010.
"Submitted to the Graduate Faculty as partial fulfillment of the requirements for the Doctor of Philosophy Degree in Biomedical Sciences." Title from title page of PDF document. "A Dissertation entitled"--at head of file. Bibliography: p. 37-41, 77-82, 102-107, 124-125, 153-160, 195-199, 221-254.
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Eynon, Elizabeth E. "Small B Cells as Antigen Presenting Cells in the Induction of Tolerance to Soluble Protein Antigens: A Dissertation." eScholarship@UMMS, 1991. https://escholarship.umassmed.edu/gsbs_diss/185.

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This thesis proposes a mechanism for the induction of peripheral tolerance to protein antigens. I have investigated the mechanism of tolerance induction to soluble protein antigens by targeting an antigen to small, resting B cells. For this purpose I have used a rabbit antibody directed at the IgD molecule found on the surface of most small, resting B cells but missing or lowered on activated B cells. Intravenous injection of normal mice with 100 μg of an ultracentrifuged Fab fragment of rabbit anti-mouse IgD (Fab anti-δ) makes these mice profoundly tolerant to challenge with nonimmune rabbit Fab (Fab NRG) fragments. This tolerance is antigen specific since treated mice make normal responses to an irrelevant antigen, chicken immunoglobulin (Ig). Fab fragments of rabbit Ig (rabbit Fab) not targeted to B cells do not induce tolerance as well as Fab anti-δ. Evidence suggests that the B cells must remain in a resting state for tolerance to be induced, since injection of F(ab)'2 anti-δ does not induce tolerance. Investigation of the mechanisms of the tolerance, by adoptive transfer, have shown that rabbit Fab specific B cell function has been impaired. The major effect however is in helper T cell function, as shown by adoptive transfer and lack of help for a hapten response. In vitro proliferation experiments show that the T cell response has not been shifted toward activation of different T cell subsets which do not help Ig production, nor is there any change in the Ig isotypes produced. Suppression does not appear to be the major cause of the helper T cell defect as shown by cell mixing experiments. This work shows that an antigen targeted to small B cells can induce tolerance to a soluble protein antigen, and suggests a role for small B cells in tolerance to self-proteins not presented in the thymus.
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Scott, Carol Elizabeth DeWeese. "Molecular modeling and experimental characterization of HLA-DQ proteins and protein/peptide complexes : correlation with insulin-dependent diabetes mellitus (IDDM) /." Thesis, Connect to this title online; UW restricted, 1997. http://hdl.handle.net/1773/8089.

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Cho, Uhn-Soo. "Structural studies of protein phosphatase 2A, simian virus 40 small t antigen, and the PhoQ sensor domain /." Thesis, Connect to this title online; UW restricted, 2007. http://hdl.handle.net/1773/5677.

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12

Spadafina, Tiziana. "Characterization of Neisseria meningitidis GNA2132 antigen, a heparin binding protein cleaved by meningococcal NalP protease." Doctoral thesis, Università degli studi di Padova, 2009. http://hdl.handle.net/11577/3427201.

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ABSTRACT GNA2132 (Genome-derived Neisseria Antigen 2132), a protein discovered by Reverse Vaccinology, is a surface-exposed lipoprotein expressed by genetically diverse Neisseria meningitidis strains. The protein induces bactericidal antibodies against most strains of meningococccus and has been included in a multivalent recombinant vaccine against N. meningitidis serogroup B. Sequence analysis of GNA2132 revealed the presence of an Arginine-rich region highly conserved among different strains. In this study we described that in meningococcus the protein is cleaved upstream from the Arginine-rich region and the C-terminal fragment is released in the culture supernatant. By genetic approaches we demonstrated that the processing of GNA2132 is strain-dependent and correlates with the expression of NalP, a phase variable autotransporter with a serine-protease activity, which has been previously shown to cleave known Neisseria virulence factors. Sequence analysis of GNA2132 protein revealed the presence of an Arginine-rich region highly conserved among N. meningitidis strains. Our functional studies revealed that this region is crucial for the function of GNA2132 since recombinant GNA2132 binds heparin through the Arginine-rich region motif.
RIASSUNTO GNA2132 (Genome-derived Neisseria Antigen 2132), una proteina scoperta tramite la Reverse Vaccinology, è una lipoproteina esposta sulla membrana esterna di Neisseria meningitidis. La proteina è in grado di indurre anticorpi battericidi contro la maggior parte dei ceppi di meningococco e per questa ragione è stata inclusa da Novartis nel vaccino contro meningococco serogruppo B. In questo studio abbiamo descritto come in meningococco GNA2132 è tagliata appena prima della regione ricca in arginine ed un frammento corrispondente alla porzione C-terminale della proteina è rilasciato nel supernatante di coltura. Tramite un approccio genetico abbiamo dimostrato che il taglio di GNA2132 è un processo ceppo-dipendente. Inoltre abbiamo dimostrato che il processamento di GNA2132 correla con l’espressione di NalP, un autotransporter ad attività serino-proteasica, la cui espressione subisce variazione di fase e che è in grado di modulare il processamento di altri fattori di virulenza di meningococco. GNA2132 contiene una regione ricca in arginine che analisi di sequenza hanno dimostrato essere altamente conservata in moltissimi ceppi di meningococco. I nostri studi sulla funzione della GNA2132 e dei frammenti generati indicano che la GNA2132 è capace di legare l’eparina e mostrano come questa regione sia fondamentale nel legame: infatti il frammento privo della regione ricca in arginine e le proteine ricombinanti (priva della zona in arginine oppure mutata nelle arginine) non mostrano avere il fenotipo di legame all’eparina.
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Michaëlsson, Jakob. "Decoding NK cell receptor specificity : functional and structural studies of MHC class 1 subcomponents /." Stockholm : [Karolinska institutets bibl.], 2002. http://diss.kib.ki.se/2002/91-7349-286-8.

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14

Anderton, Stephen M. "A study of murine T lymphocyte responses to Streptococcus pyogenes." Thesis, University of Newcastle Upon Tyne, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.287332.

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15

Forsman, Alma. "The Epstein-Barr virus nuclear antigens 1 & 5 : study of virus-host cellular protein interactions /." Göteborg : Institute of Biomedicine, Department of Clinical Chemistry and Transfusion Medicine, 2009. http://hdl.handle.net/2077/21357.

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16

Lilyestrom, Wayne. "The structure of the complex containing the oncoprotein SV40 large tumour antigen bound to the p53 tumor suppressor /." Connect to full text via ProQuest. IP filtered, 2006.

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Thesis (Ph.D. in Biochemistry) -- University of Colorado, 2006.
Typescript. Includes bibliographical references (leaves 69-83). Free to UCDHSC affiliates. Online version available via ProQuest Digital Dissertations;
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Clarke, Christopher John. "Antigen presentation to mucosal surfaces : the influence of liposome entrapment and cholera toxin on the immune response to fed protein antigens." Thesis, University of Bristol, 1989. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.330040.

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18

Ramsay, Lesley Anne. "Humoral responses to acetaldehyde-modified protein antigens." Thesis, Anglia Ruskin University, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.286770.

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19

Seabrook, Richard N. "An immunochemical study of B. pertussis antigens." Thesis, Queen Mary, University of London, 1990. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.279600.

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20

Nguyen, Teresa. "Application of a Whole Genome Approach to the High Throughput Discovery of Novel Diagnostic Antigens for Brucella abortus." Thesis, Université d'Ottawa / University of Ottawa, 2020. http://hdl.handle.net/10393/40715.

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Brucella abortus is the etiological agent of bovine brucellosis, a zoonotic disease that can be transmitted to humans through direct contact with infected animals or consumption of contaminated food products. Current serological tests used to identify infected animals rely on the detection of antibodies to O-antigens of the smooth lipopolysaccharide (sLPS) of B. abortus. Due to the presence of structurally similar O-antigens of bacteria such as E. coli O:157 and Yersinia enterocolitica O:9, these tests can produce false-positive results, which requiring alternative protein target antigens. We hypothesize that through comparative genomics and bioinformatics analysis of all ORFs within the B. abortus genome, followed by profiling of the humoral immune responses to surface and extracellular proteins, novel protein antigens with diagnostic potential may be discovered. In this study, the genomes of thirteen strains were analyzed using a subcellular localization prediction database (PSORTb) to identify proteins in the outer membrane and extracellular space. A total of 100 ORFs coding for such proteins including known immunogenic proteins reported in literature were identified and selected for recombinant protein expression using high-throughput in vivo cloning and in vitro transcription/translation strategies. The in vitro expression of 67 of these candidates has been successfully demonstrated. These recombinant B. abortus proteins were subsequently probed with E. coli pre-adsorbed sera from infected animals for the identification of immunoreactive protein antigens. Ten unique candidates were demonstrated to be antigenic and have the potential for diagnostic applications. This study illustrates a unique, high throughput strategy to express and screen proteins of a bacterial pathogen for novel diagnostic antigen discovery.
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21

Pickering, Maureen Gabrielle. "Immune responses to ingested protein antigens in mice." Thesis, University of Edinburgh, 1985. http://hdl.handle.net/1842/19235.

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22

Boulton-Jones, John Robert. "Oral tolerance to soluble protein antigens in humans." Thesis, University of Edinburgh, 2001. http://hdl.handle.net/1842/22766.

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In this thesis, a protocol to demonstrate oral tolerance is described. Keyhole limpet haemocyanin (KLH), a neo-antigen, was used to demonstrate low dose tolerance. A control group was immunised with KLH and the subsequent immune response was assessed by delayed type hypersensitivity responses, in vitro lymphocyte proliferation to KLH and anti-KLH IgG and IgA production. Another group of volunteers was pre-fed a course of KLH prior to receiving the same immunisation schedule. The same measures of the immune response used in the control group were assessed and any differences were attributed to oral tolerance. A third group of volunteers was immunised with ovalbumin (OVA) and the immune response was measured. OVA is a common dietary protein and therefore was used to assess oral tolerance to extended courses of feeding. The group fed 50mg of KLH for 10 days demonstrated reduced DTH responses but without significant differences in in vitro lymphocyte proliferation and priming of anti-KLH IgG production. These changes demonstrated that oral tolerance can occur in the T cell compartment in humans after a short course of antigen feeding. In contrast, those volunteers immunised with OVA showed no in vitro lymphocyte proliferation or DTH responses. Anti-OVA IgA and IgA were detectable in low levels prior to immunisation but there was no increase in humoral response after immunisation. These results suggest that oral tolerance is more pronounced to an antigen that has been encountered over prolonged periods. The nature of the tolerance to KLH is not known. It may be the result of the induction of immunoregulatory cells or the induction of clonal anergy. Experiments were designed to test for the presence of immunoregulatory cells induced by feeding KLH, but no positive results were obtained. The mechanisms maintaining tolerance to OVA are likewise unknown. Attempts to demonstrate clonal anergy by reserving tolerance with IL-2 failed.
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23

LOMBARDI, BENEDETTA. "Staphylococcal antigens characterization through a protein interaction approach." Doctoral thesis, Università del Piemonte Orientale, 2014. http://hdl.handle.net/11579/46757.

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24

NILO, ALBERTO. "Exploring carbohydrates antigens conjugated to pathogenrelated protein carriers." Doctoral thesis, Università del Piemonte Orientale, 2015. http://hdl.handle.net/11579/115558.

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25

Hauser, Thomas. "Definition immunogener Antigene bei der Akuten Myeloischen Leukämie mittels Serologischem Screening von cDNA-Expressionsbanken." [S.l. : s.n.], 2006. http://nbn-resolving.de/urn:nbn:de:bsz:289-vts-56608.

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26

Peel, Mary. "A study of the protein antigens of Listeria monocytogenes." Thesis, University of Edinburgh, 1988. http://hdl.handle.net/1842/29939.

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27

Weiss, Timo. "Funktionelle Charakterisierung des granulozytären Antigens NB1 (CD177)." Giessen : VVB Laufersweiler, 2008. http://d-nb.info/991426673/04.

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28

Leishman, Andrew James. "Intervention in established immune responses by feeding protein antigens." Thesis, University of Glasgow, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.301603.

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29

Willhite, David Grant. "Purification, identification, and partial characterization of proteins associated with the adaptive immune response to soluble protein antigen in the American cockroach." Cincinnati, Ohio : University of Cincinnati, 2001. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=ucin1005252852.

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30

ALVES, JANAINA B. "Aspectos da resposta imune frente a antigenos proteicos irradiados com sup(60)Co." reponame:Repositório Institucional do IPEN, 2004. http://repositorio.ipen.br:8080/xmlui/handle/123456789/11183.

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Dissertacao (Mestrado)
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Instituto de Pesquisas Energeticas e Nucleares - IPEN/CNEN-SP
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31

Willis, Dean. "The role of heat shock proteins in models of acute inflammation." Thesis, Queen Mary, University of London, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.265943.

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32

Gall, Jason G. D. "The role of adenovirus fiber and hexon proteins in virus-host interactions /." Access full-text from WCMC:, 1998. http://proquest.umi.com/pqdweb?did=1432807591&sid=9&Fmt=2&clientId=8424&RQT=309&VName=PQD.

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33

Nassar, M. I. A. "Immunohistochemical studies of clotting factors in renal disease." Thesis, University of Southampton, 1987. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.383291.

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34

Wang, Yibing. "Study of cysteines in the stalk region of CD3 proteins : evolutionarily conserved residues critical for T cell development and function /." Connect to full text via ProQuest. Limited to UCD Anschutz Medical Campus, 2008.

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Thesis (Ph.D. in Immunology) -- University of Colorado Denver, 2008.
Typescript. Includes bibliographical references (leaves 138-153). Free to UCD Anschutz Medical Campus. Online version available via ProQuest Digital Dissertations;
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35

Gannon, Julian Vaughan. "Interactions between SV40 large T antigen and host proteins." Thesis, Institute of Cancer Research (University Of London), 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.314829.

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36

Herberg, Jethro Adam. "Antigen presenting genes and proteins of the human MHC." Thesis, King's College London (University of London), 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.298847.

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37

Gangloff, Sophie. "Les adhesines des bacteries du groupe mutans : cartographie antigenique d'une proteine de surface de streptococcus mutans." Strasbourg 1, 1991. http://www.theses.fr/1991STR15070.

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38

Maloy, Kevin Joseph. "The basis of the oral and parenteral adjuvant properties of immune stimulating complexes (ISCOMS)." Thesis, University of Glasgow, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.321045.

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39

Grazette, Affif. "Molecular characterisation of squamous cell carcinoma antigen recognised by T-cells 3, an adaptor protein of ubiquitin specific protease 15." Thesis, University of Nottingham, 2016. http://eprints.nottingham.ac.uk/33081/.

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The deubiquitinating enzyme USP15, a member of the USP family, reverses the process of ubiquitination thereby altering the fate of a plethora of substrates. As such, USP15 has been implicated in a numerous important cellular pathways including cell cycle progression, transcriptional modification and DNA damage repair. The spliceosomal subunit recycling protein SART3 has been shown to bind to USP15 enhancing its deubiquitination of histone H2B thereby providing histone dimers for reassembly during subsequent rounds of transcription and splicing. Furthermore SART3 has also been shown to bind to USP4, a close homologue of USP15. In addition to this histone chaperone activity, SART3 primarily functions by mediating the re-annealing of the U4 and U6 snRNPs to facilitate consequent rounds of splicing in addition to its implication in several disease states including numerous cancer types and HIV through interactions with various cellular proteins. In this thesis ITC, ESI-MS, analytical size exclusion, X-ray crystallography, SAXS and pull-down assays are used to characterise the interaction between USP15 and SART3, solve the structure of the N-terminus of SART3 and identify novel binding partners of the USP15-SART3 complex. The structure of SART3’s N-terminus (residues 96-574) has been solved to 3.04Å, revealing that it is a homodimer comprised of a series of anti-parallel α-helices that form a shape reminiscent of a bowtie. This dimeric arrangement is retained in solution and can bind two molecules of USP15DU. The DU-finger of USP15 plays a pivotal role in co-ordinating the binding interaction with SART3, as small changes to this region can affect the nanomolar affinity interaction between USP15 and SART3. In addition the USP15-SART3 complex appears to interact with several cellular proteins which could have significant impact in several disease states.
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40

Jarmin, Sarah Jane. "Investigation of protein tyrosine phosphatases in antigen receptor signalling." Thesis, University of Cambridge, 2003. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.619653.

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41

Guo, Beichu. "Interaction of PKCbeta with CARMA1 mediates B cell receptor-induced NF-kappaB activation /." Thesis, Connect to this title online; UW restricted, 2003. http://hdl.handle.net/1773/8348.

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42

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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43

AZZOUZ, NAHID. "Etude d'une glycosyl-phosphatidylinositol proteine : antigene de surface 156g de paramecium primaurelia." Paris 6, 1991. http://www.theses.fr/1991PA066402.

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Le protozoaire, paramecium aurelia, possede un repertoire d'antigenes de surface dont l'expression, mutuellement exclusive, est sous la dependance de parametres externes. Il s'agit d'isoproteines de hauts poids moleculaire, environ 300 kd, riches en threonine, serine et cysteine, sans fonction thiol libre. Il a ete etabli par une approche indirecte que ces proteines sont ancrees dans la membrane plasmique par un glycosyl phosphatidyl-inositol (gpi) et qu'il existait une phospholipase c endogene (pi-plc) capable de solubiliser ces molecules par clivage du diacylglycerol. Notre travail a porte sur l'antigene de surface g de la souche 156 de p. Primaurelia, qui est exprime a moyenne temperature et dont le gene entierement sequence a revele l'existence d'une structure periodique. Nous avons mis au point un protocole de purification en masse de la proteine 156g sous la forme membranaire et demontre par marquages biosynthetiques que les constituants caracteristiques de l'ancre gpi, tels que l'ethanolamine, le myo-inositol, le phosphate et un acide gras, en l'occurrence le myristate, etaient presents dans la proteine purifiee. Un peptide hydrophobe portant l'ancre gpi a ete purifie par hplc en phase reverse apres digestion de la proteine par la pronase. Nous avons purifie la forme soluble de l'antigene 156g a partir de la forme membranaire apres action d'une pi-plc exogene bacterienne, dans le but de comparer ces deux formes. Nous avons montre que seule la forme membranaire pouvait etre incorporee dans des liposomes et dans les fantomes de globules rouges. Par des etudes physico-chimiques (chromatographie d'exclusion, dichroisme circulaire, ultracentrifugation analytique) nous avons montre que les deux formes d'antigene, la forme membranaire et la forme soluble se comportaient de la meme facon. Ceci suggere donc que les structures secondaires et tertiaires demeurent essentiellement inchangees dans la forme soluble obtenue par clivage enzymatique de l'ancre lipidique. Une comparaison des mesures hydrodynamiques et des donnees obtenues par diffusion des rayons x indique qu'il y a une distribution asymetrique de la masse. La microscopie electronique de la particule montre qu'il s'agit bien d'une particule allongee
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44

Chan, S. W. "Characterisation of a novel heat shock protein-enriched multivalent Streptococcus pneumoniae protein antigen vaccine." Thesis, University College London (University of London), 2016. http://discovery.ucl.ac.uk/1524189/.

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Current vaccination against Streptococcus pneumoniae uses vaccines based on capsular polysaccharides from a limited number of serotypes, which has led to replacement by non-vaccine serotypes. An alternative approach based on multiple protein antigens enriched with and bound to highly conserved heat-shock proteins (Hsps) could provide multivalent protection, thereby preventing the problem of serotype replacement. The work in this thesis using proteomic techniques shows that the Hsp vaccine preparation resulted in a product that was enriched with Hsps and contained several known important antigens. The Hsp vaccine induced robust antibody responses in rodents and serum from vaccinated animals contained cross-reactive antibodies against multiple serotypes, including non-vaccine serotypes. Homologous and heterologous strains of S. pneumoniae were opsonised with Hsp vaccine-induced IgG. In mouse models of active vaccination significantly protected against pneumonia infection whilst passive vaccination of rabbit serum significantly protected against sepsis caused by both the homologous and a heterologous S. pneumoniae strain. These data suggest that the Hsp vaccine has the potential to provide serotype-independent protection against S. pneumoniae. Further work in this thesis also showed that protective immunity induced by prior exposure to S. pneumoniae, either by intraperitoneal injection of killed bacteria or by colonisation with live bacteria, targeted protein antigens and was enhanced by exposure to multiple S. pneumoniae strains. In particular, immunisation with multiple strains induced broader cross-reactive immunity to heterologous strains than exposure to only a single strain. This offers insight into how naturally acquired immunity to multiple S. pneumoniae strains develops in humans, and suggests that vaccines derived from multiple strains will provide stronger heterologous protection.
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45

Kohli, Évelyne. "Etude des sites antigeniques de la proteine majeure de capside interne du rotavirus et application au diagnostic." Dijon, 1990. http://www.theses.fr/1990DIJO0001.

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46

Yan, Wei Xing. "Target antigen in chronic inflammatory demyelinating polyradiculoneuropathy." Thesis, The University of Sydney, 2001. https://hdl.handle.net/2123/27718.

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The primary purposes of this study were to investigate humoral factors in chronic inflammatory demyelinating polyradiculoneuropathy (CIDP), to determine pathogenicity of the factors found, and to detect possible target antigens in CIDP. Sera from 22 CIDP patients of whom 12 responded to plasma exchange therapy and 10 needed other therapy, were tested by means of enzyme-linked immunosorbent assay, immunofluorescence and Western blot techniques. Anti-whole myelin, anti-myelin glycoproteins (e.g. P0, P2, PMP22 and myelin associated glycoprotein), anti-myelin glycolipids (e.g. GMl, GM2, Gle, GDla, Gle, Gal-C and Gal-S) antibodies and complement reactivity were detected. Using systemic administration after opening the blood-nerve barrier (BNB) and intraneural injection, Lewis rats were assessed electrophysiologically, morphologically and clinically before and after administrated CIDP serum. Methods of antibody purification and protein sequencing were also applied. The results revealed that one or more antibodies to whole peripheral nerve myelin, myelin glycolipids (GMl, Gle, Qle, GM2 and Gal-S) and myelin glycoproteins (P0) and complement (C3) reactivity were found in 16 (73%) of 22 CIDP sera. Six had anti—P0 IgG antibody, four of which had antimyelin IgG antibody and C3 reactivity. Five had anti-myelin IgM antibody, of which four were directed against gangliosides and two showed C3 reactivity. Five had anti-ganglioside antibody, of which two showed C3 reactivity. One had C3 reactivity alone. Five had neither anti-PO, anti-myelin or anti— ganglioside antibody, nor C3 reactivity. No sera had detectable antibodies to myelin associated glycoprotein, PMP22, P2, GDla or Gal-C. Pathogenicity to peripheral nerve was demonstrated in four of six sera which contained IgG anti-P0 antibody and C3 reactivity. Conduction block and demyelination were induced by intraneural injection of these four sera and their IgG, and by systemic transfer when the BNB had been opened by activated T cells. The specificity of the antigen-antibody reaction was displayed by the binding of F(ab)2 to the P0 protein. Loss of pathogenicity was showed following absorption by P0 but not by P2. Protein sequencing showed that the target antigen is a 30kDa peripheral nervous system myelin protein, the P0 protein.
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47

BOULANGER, FLORENCE. "Toxoplasma gondii : identification, localisation et quantification de quatre antigenes immunodominants : conformation moleculaire et relation structure-activite de l'antigene majeur p30." Reims, 1991. http://www.theses.fr/1991REIMM204.

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48

Carson, Richard Thomas. "Immunoglobulin G subclass responses to bacterial protein antigens in man." Thesis, University of Newcastle Upon Tyne, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.384823.

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49

Shand, Geoffrey H. "Antibiotic resistance and outer membrane protein antigens of Pseudomonas aeruginosa." Thesis, Aston University, 1985. http://publications.aston.ac.uk/12475/.

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50

Weiland, Malin. "Immunodominant proteins in Giardia lamblia /." Stockholm, 2004. http://diss.kib.ki.se/2004/91-7140-158-X/.

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