prikaz prve stranice dokumenta Characterization of antibodies specific for MCMV proteins MCK-2 and gO
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master's thesis
Characterization of antibodies specific for MCMV proteins MCK-2 and gO
2017. urn:nbn:hr:193:452978
Jerak, Josipa
University of Rijeka
Faculty of Biotechnology and Drug Development

Cite this document

Jerak, J. (2017). Characterization of antibodies specific for MCMV proteins MCK-2 and gO (Master's thesis). Rijeka: University of Rijeka. Retrieved from https://urn.nsk.hr/urn:nbn:hr:193:452978

Jerak, Josipa. "Characterization of antibodies specific for MCMV proteins MCK-2 and gO." Master's thesis, University of Rijeka, 2017. https://urn.nsk.hr/urn:nbn:hr:193:452978

Jerak, Josipa. "Characterization of antibodies specific for MCMV proteins MCK-2 and gO." Master's thesis, University of Rijeka, 2017. https://urn.nsk.hr/urn:nbn:hr:193:452978

Jerak, J. (2017). 'Characterization of antibodies specific for MCMV proteins MCK-2 and gO', Master's thesis, University of Rijeka, accessed 16 November 2024, https://urn.nsk.hr/urn:nbn:hr:193:452978

Jerak J. Characterization of antibodies specific for MCMV proteins MCK-2 and gO [Master's thesis]. Rijeka: University of Rijeka; 2017 [cited 2024 November 16] Available at: https://urn.nsk.hr/urn:nbn:hr:193:452978

J. Jerak, "Characterization of antibodies specific for MCMV proteins MCK-2 and gO", Master's thesis, University of Rijeka, Rijeka, 2017. Available at: https://urn.nsk.hr/urn:nbn:hr:193:452978

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Metadata
TitleCharacterization of antibodies specific for MCMV proteins MCK-2 and gO
Title (croatian)Karakterizacija protutijela specifičnih za MCMV protein MCK-2 i gO
AuthorJosipa Jerak
MentorIlija Brizić (mentor)
MentorStipan Jonjić (mentor)
Committee memberIvana Munitić (predsjednik povjerenstva)
Committee memberStipan Jonjić (član povjerenstva)
Committee memberIgor Jurak (član povjerenstva)
GranterUniversity of Rijeka
(Faculty of Biotechnology and Drug Development)
Rijeka
Defense date and country2017-10-30, Croatia
Scientific / art field,
discipline and subdiscipline		BIOTECHNICAL SCIENCES
Biotechnology
Abstract
Cytomegalovirus (CMV) is β-herpesvirus, which establishes lifelong latent infection in host. CMV is transmitted through bodily fluids and represents the most frequent congenital infection in humans. Infection of healthy individuals with human CMV (HCMV) is usually asymptomatic, however, it can cause severe disease in immunocompromised individuals, as well as in infants infected in the prenatal period. CMV has broad tropism, infecting majority of cell types and organs. Viral glycoproteins mediate fusion with host cell membranes and entry of virus into target cells. Glycoprotein complexes gH/gL are determining virus specificity for infection of distinct cell types. Due to the species-specific nature of CMV, mouse CMV (MCMV) has been widely used as a model to study HCMV infection. Two gH/gL complexes have been identified in MCMV, gH/gL/gO and gH/gL/MCK-2. However, the biology and role of these complexes is still incompletely defined, in part due to the lack of antibodies specific for these proteins. Here I characterized new antibodies specific for gO and MCK-2 using flow cytometry, Western blot and enzyme-linked immunosorbent assay. Antibodies were generated by immunizing mice with proteins produced in eukaryotic cells and subsequent fusion of B cells with myeloma cells. Obtained hybridomas were preliminary screened by ELISA and positive antibodies were further tested in Western blot and flow cytometry. In total, seven out of fifty-one tested MCK-2 antibodies were specific for MCK-2 in flow cytometry. Five of them were positive in Western blot as well. Furthermore, five out of twelve tested gO antibodies specific for gO were identified by flow cytometry, and additional four by Western blot. Identified antibodies will be used in future studies of MCMV gH/gL complexes.
Abstract (croatian)
Citomegalovirus (CMV) je β-herpesvirus koji u domaćinu uspostavlja doživotnu latentnu infekciju. CMV se prenosi tjelesnim tekućinama i predstavlja najčešću prirođenu infekciju u ljudi. Infekcija ljudskim CMV-om (eng. Human CMV, HCMV) je kod zdravih ljudi obično asimptomatska, međutim kod osoba sa oslabljenim imunološkim sustavom te djece zaražene u prenatalnom razdoblju može izazvati kliničku manifestaciju bolesti. CMV inficira većinu staničnih tipova i posljedično većinu organa. Fuzija virusne i stanične membrane te ulazak virusa u stanicu su posredovani virusnim glikoproteinima. Specifičnost virusa za ulazak u različite tipove stanica je određena glikoproteinskom kompleksima gH/gL. Budući da je HCMV specifičan za vrstu, mišji CMV (eng. mouse CMV, MCMV) se koristi kao model za istraživanje infekcije ljudskim CMV-om. Dva glikoproteinska kompleksa gH/gL su karakterizirana u mišjem CMV-u: gH/gL/gO i gH/gL/MCK-2. Biološke funkcije i uloge ovih kompleksa su nepotpuno razjašnjene, djelomično zbog nedostatka specifičnih protutijela. U ovom radu sam karakterizirala protutijela specifična za proteine gO i MCK-2 koristeći protočnu citometriju, Western blot i enzimski vezani imunosorbentni test (ELISA). Protutijela su dobivena imunizacijom miševa proteinima proizvedenim u eukariotskim stanicama nakon čega je uslijedila fuzija B limfocita sa stanicama mijeloma. Dobiveni hibridomi preliminarno su analizirani uz pomoć ELISA-e te su pozitivna protutijela dalje analizirana u Western blotu i protočnoj citometriji. Sedam od ukupno pedeset dva testirana MCK-2 protutijela je bilo specifično za MCK-2 u protočnoj citometriji, a njih pet je također bilo pozitivno u Western blotu. Nadalje, pet od ukupno dvanaest testiranih gO protutijela bilo je specifično za gO u protočnoj citometriji, a dodatnih četiri u Western blotu. Identificirana protutijela će biti korištena u daljnim istraživanjima gH/gL kompleksa MCMV-a.
Keywords
Keywords (croatian)
Languageenglish
URN:NBNurn:nbn:hr:193:452978
Study programmeTitle: Biotechnology in medicine
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra biotehnologije u medicini (magistar/magistra biotehnologije u medicini)
Type of resourceText
File originBorn digital
Access conditionsAccess restricted to students and staff of home institution
Terms of useLicence In copyright icon
Created on2018-01-15 09:49:23