National Repository of Grey Literature 27 records found  1 - 10nextend  jump to record: Search took 0.01 seconds. 
Synthesis, characterization and study of the properties of functional antibody conjugates
Stránská, Andrea ; Hrubý, Martin (advisor) ; John, Jan (referee)
Radioimmunotherapy combines biological treatment with the use of highly effective radioactive radiation, which allows for more effective and gentle cancer therapy. This bachelor thesis focuses on the use of the therapeutic radioisotope 161Tb, which is characterized by the emission of low-energy electrons and gamma rays that allow monitoring of therapy and targeting of cancer cells, metastases and whole tumors. This theranostic approach offers increased treatment efficacy, especially in the early stages of the disease. This thesis focuses on a model study of rituximab monoclonal antibody conjugates that have been fluorescently labelled using a fluorescein tag. Covalent binding between the immunotherapeutic conjugate and the fluorescent label was accomplished using free amino groups of lysines and bioorthogonal chemistry. A biologically active antibody conjugate with a fluorescent label was developed that allowed direct monitoring of the conjugate binding to CD20-expressing cells using confocal microscopy. Subsequently, a radioactive conjugate of rituximab, modified with the p-SCN-Bn-CHX-A"-DTPA chelator and radiolabeled with the 161 Tb radioisotope, was tested. The biological activity of the radioimmunoconjugate was determined by in vitro experiments. Biodistribution of the radioimmunoconjugate was...
Effect of porcine reproductive and respiratory syndrome virus (PRRSV) on the adaptive immune system.
Šinkora, Šimon ; Štěpánová, Kateřina (advisor) ; Toman, Miroslav (referee)
The porcine reproductive and respiratory syndrome virus (PRRSV) causes large economic losses in pork production because of its ability to persist and spread in herds despite vaccination. An essential feature of the infection is a strong but ineffective antibody response with limited immunological memory. The ability of the virus to dysregulate the host immune system is explained by various mechanisms, including antibody-dependent enhancement of infection, polyclonal activation of B cells by the superantigen effect, the presence of immunodominant epitopes, inhibition of nonspecific immunity, or a different role of cytotoxic T cells. Recent evidence suggests that PRRSV affects the nascent T cell repertoire in the thymus and induces tolerance to viral epitopes that are crucial for virus neutralization. Specific T helper cells are absent and cannot initiate an adequate B cell response to produce high-affinity virus-neutralizing antibodies. However, the remaining T helper cells are not affected and help to produce anti-PRRSV antibodies, which are ineffective and cannot eliminate the virus. The cytotoxic T cells needed for the elimination of infected cells are also affected. This bachelor thesis describes the various features of PRRSV infection and compares the known information on the immune response...
Cancer Immunotherapy exploiting engineered antibody fragments against prostate-specific membrane antigen
Das, Gargi ; Bařinka, Cyril (advisor) ; Vaněk, Ondřej (referee) ; Ormsby, Tereza (referee)
Prostate cancer (PCa) remains a leading cause of male cancer-related mortality, necessitating thus the development of novel therapeutic approaches as conventional treatments have limited efficacy. Prostate-specific membrane antigen (PSMA) is an established biomarker for both imaging and therapy of PCa, as it is highly upregulated in neoplastic PCa tissues and metastatic castration- resistant prostate cancer. Consequently, immunological targeting of PSMA has gained significant attention as a therapeutic platform for the management of the disease. The thesis is focused on engineering of antibody fragments and fusion proteins derived from the high affinity anti-PSMA 5D3 monoclonal antibody that can be used as immune cell engagers to target and eliminate PSMA-positive cells. To this end, we engineered 5D3 single chain variable fragments (scFv) that were subsequently fused to anti-CD3 scFv and CP33 sequences, creating thus immune cell engagers targeting T-cells (BiTE) and monocytes (5D3-CP33), respectively. The engagers were expressed in insect cells, purified to homogeneity and their biophysical and functional characteristics evaluated using size exclusion chromatography, differential scanning fluorimetry, ELISA and flow cytometry. Ensuing cell-based assays revealed that both BiTE and 5D3-CP33 can...
Predicting local structural properties from antibody sequence
Beňo, Roman ; Příhoda, David (advisor) ; Hoksza, David (referee)
Predicting local structural properties of antibodies at residual level is vital for detecting the presence of post-translational modifications (PTMs), which often induce structural change in the antibody, negatively impact its shelf-life and possibly lead to the loss of the therapeutic potential. In this work, we predict relative solvent accessibility (RSA) of individual residues. This property is, alongside with the type of amino acid in question, the key indicator for presence of methionine oxidations and other types of PTMs. Due to the conservation of the antibody structure, we identified that different classes of prediction methods yield almost interchangeable results - total mean absolute error (MAE) of 5.64 RSA percentage units measured for the best performing machine learning pipeline compared to the 5.96 measured for the best performing statistical pipeline. The significant prediction quality improvement observed within comparison to the random prediction method with MAE of 35.996 may be as well attributed to the sequence conservancy. In CDR regions, RSA values are harder to predict. Although the range of methods and procedures employed throughout this work is by far not able to yield complex structure predictions, it might constitute a modular, high-throughput tool to support one's choices when...
Implementation and optimization of ELISA method
KOPAČKOVÁ, Tereza
Enzyme-Linked ImmunoSorbent Assay or ELISA is a method that has a wide application, especially in immunology. Antigens and antibodies can be detected by the method. The basic principle of the method is the reaction of antigen and antibody and the subsequent formation of an immunocomplex. The immunocomplex is detected by a conjugate that causes a color change of the substrate in the presence of antigen or antibody in the test sample. The ELISA test is a simple method important fot the diagnosis of various diseases or laboratory research. This method is a special type of enzyme immunoassay. The bachelor thesis is focused on the general implementation and optimization of the ELISA method. The aim of the theoretical part of the work is a general introduction to immunological methods. The division of individual types of ELISA tests, including the principles and use in practice, is elaborated in detail. In the practical part, the ELISA method is introduced and optimized using a DYNAREAD photometer for reading 96well microtiter plates and AlaDYN software. In this way, a total of 8 test samples of sera are examined and subsequently the obtained data are processed. The test measurement was performed using the QuatiVac ELISA test kit and human IgG antibodies against SARS-CoV-2 in serum were determined. The peaks of SARS-CoV-2 spike proteins expresses recombinantly in the human cell line HEK 293 were used as antigens. The measurement was performed using a DYNAREAD instrument and data processing was performer using AlaDYN software. 6 calibrators, a positive control, a negative control and then individual serum samples were measured. The measurement results are 5 serum samples in which IgG class anti-SARS-CoV-2 antibodies were detected and 3 samples in which no antibodies were detected.
Úloha osy PD-1/PD-L1 při infekci \kur{Borrelia burgdorferi} u myší
PALOUNKOVÁ, Anna
Borrelia burgdorferi, the causative agent of Lyme disease, induces upregulation of inhibitory immune checkpoint PD-L1 in mice. We studied if the blockade of PD-1/PD-L1 axis by neutralizing antibodies influences the proliferation of T lymphocytes and cytokine milieu in imunological synapsis between murine dendritic cells and T cells in vitro.
Diagnostic of immunohistochemistry in pathology
HRDINOVÁ, Andrea
The bachelor thesis deals with the use of immunohistochemical methods in the diagnosis of neoplasms in pathology. Describes the use of antibody spectrum at the Department of Pathology, Český Krumlov Hospital, a.s. Immunohistochemical methods are nowadays an important method used in pathology for tumor typing and metastasis for the determination of primary tumors. They complement the standard staining methods for basic diagnostics such as Hematoxylin-Eosin and Van-Gieson, which are less specific, these methods mainly serve to differentiate the basic cellular components and tissue components of the preparation. Immunohistochemistry methods are based on the special antibody affinity principle and are divided into direct and indirect methods. For a direct method, an antibody labeled with, for example, an enzyme is used to detect the antigen and, upon binding, induces a color change observed under the microscope. Indirect method (two-stage, three-stage) is a more sensitive method, based on the principle of an unlabeled primary antibody to which a conjugated secondary antibody binds to an enzyme or biotin. In the three-stage, we use the affinity of avidin and biotin to form the avidin-biotin-enzyme complex (eg. peroxidase), which is shown and highlighted. In more detail, the thesis deals with the diagnosis of breast and sentinel lymph node tumors, both in the theoretical and practical part, and the incidence of these tumors since the introduction of the immunohistochemical method at the Department of Pathology of the Český Krumlov Hospital. Last but not least, a case report of individual interesting cases is described.
Current approaches in the development of vaccines against infectious viral diseases
Vargová, Soňa ; Malý, Petr (advisor) ; Osička, Radim (referee)
Vaccination remains one of the most successful biomedical interventions for preventing viral diseases. While early vaccines were developed by attenuating the infectious agent in cell cultures or by inactivation, new delivery platforms are on the rise thanks to the advent of genetic engineering. The COVID-19 pandemic stimulated the rapid adoption and a massive deployment of these platforms. Viral vector vaccines elicit antigen expression within cells and induce a robust cytotoxic T cell response, unlike protein subunit vaccines conferring mainly humoral immunity. mRNA vaccines also deliver the antigen inside the cells while offering more manageable and faster manufacturing possibilities. Unlike DNA-based vaccines, mRNA does not enter the nucleus, and thus, the probability of disrupting gene expression in the recipient cell is diminished. This thesis aims to offer an overview of current approaches in vaccinology and discuss the various platforms in use. The thesis will also present recent advances in the development of prophylactic vaccines against infections with human immunodeficiency virus-1 (HIV-1) and hepatitis C virus (HCV) and also will focus on a recently proposed strategy for vaccine development based on non-cognate ligands mimicking epitopes recognised by broadly neutralising antibodies...
Immunity in hosts repeatedly exposed to sand fies and the effect on pathogen transmission
Pohanková, Lucia ; Kolářová, Iva (advisor) ; Fialová, Anna (referee)
6 Abstrakt During the feeding of infected sand flies are inoculate into the host also sand fly saliva, which can strongly modulate the response of the immune system. If the host is naive, the course of infection is usually worse. In cutaneous leishmaniasis, the lesions developed early, are more destructive and persist longer. The hosts living in endemic areas of leishamniasis and their vector hosts are often exposed to feeding uninfected sand flies. For host are the saliva antigenic and induces specific cellular and antibody responses. This responses induce the protection against leishmania infection and differ for different hosts, attempts were made most frequently in murine and canine models. In humans, as hosts is difficult to monitor developments leishmania infection after previous exposure, because in humans mainly monitors the levels of antibodies, by which we can determine the degree of sand fly bites and the risk of transmission of leishmaniasis. Keywords: Lutzomyia, Phlebotomus, DTH, antibody, IgG,
Diagnostics and management of aloimunization in pregnancy
Kučerová, Rebecca ; Hornová, Markéta (advisor) ; Žižka, Zdeněk (referee)
The bachelor thesis focuses on the issue of maternal alloimmunization in pregnancy and deals with diagnostic and management of the alloimmunization. This complication would occur in approximately 2000 women in the Czech Republic per a year without a treatment. A pregnant woman is at risk if she is Rh(D) negative and the fetus inherits Rh(D) positivity from father. The development of alloimmunization can be prevented by early preventive dosage of anti-D globulin. Despite a systematically organized system of prophylaxis, a small number of pregnant women still experience immunization. Fetuses are at risk of developing anemia in the uterus and than after birth by developing hyperbilirubinemia. In the most severe cases of intrauterine anemia, the solution is intraumbilical transfusion of erythrocytes. The aim of this study was to summarize the outcomes of IUT in retrospect. The quantitative practical part is completed with 2 model study cases of hydrops fetalis. The quantitative part of the research analyzes the results of all IUTs performed in the period from 1991 to 2021 in the Center of Fetal Medicine of the Gynecology and Obstetrics Clinic of the General Hospital and the First Medical Faculty of Charles University in Prague. Using tables and graphs, the data were then analyzed and described in...

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