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Naslov:TRIM28 and [beta]-actin identified via nanobody-based reverse proteomics approach as possible human glioblastoma biomarkers
Avtorji:ID Jovchevska, Ivana (Avtor)
ID Šamec, Neja (Avtor)
ID Kočevar Britovšek, Nina (Avtor)
ID Cesselli, Daniela (Avtor)
ID Podergajs, Neža (Avtor)
ID Limbaeck Stanic, Clara (Avtor)
ID Myers, Michael P. (Avtor)
ID Muyldermans, Serge (Avtor)
ID Hassanzadeh Ghassabeh, Gholamreza (Avtor)
ID Motaln, Helena (Avtor)
ID Ruaro, Maria Elisabetta (Avtor)
ID Bourkoula, Evgenia (Avtor)
ID Lah Turnšek, Tamara (Avtor)
ID Komel, Radovan (Avtor)
Datoteke:URL URL - Izvorni URL, za dostop obiščite http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0113688
 
.pdf PDF - Predstavitvena datoteka, prenos (652,32 KB)
MD5: AA68A2E044D5298AA9A30E5A88CB9306
 
URL URL - Izvorni URL, za dostop obiščite https://doi.org/10.1371/journal.pone.0113688
 
Jezik:Angleški jezik
Tipologija:1.01 - Izvirni znanstveni članek
Organizacija:Logo NIB - Nacionalni inštitut za biologijo
Povzetek:Malignant gliomas are among the rarest brain tumours, and they have the worst prognosis. Grade IV astrocytoma, known as glioblastoma multiforme (GBM), is a highly lethal disease where the standard therapies of surgery, followed by radiation and chemotherapy, cannot significantly prolong the life expectancy of the patients. Tumour recurrence shows more aggressive form compared to the primary tumour, and results in patient survival from 12 to 15 months only. Although still controversial, the cancer stem cell hypothesis postulates that cancer stem cells are responsible for early relapse of the disease after surgical intervention due to their high resistance to therapy. Alternative strategies for GBM therapy are thus urgently needed. Nanobodies are single-domain antigen-binding fragments of heavy-chain antibodies, and together with classical antibodies, they are part of the camelid immune system. Nanobodies are small and stable, and they share a high degree of sequence identity to the human heavy chain variable domain, and these characteristics offer them advantages over classical antibodies or antibody fragments. We first immunised an alpaca with a human GBM stem-like cell line prepared from primary GBM cultures. Next, a nanobody library was constructed in a phage-display vector. Using nanobody phage-display technology, we selected specific GBM stem-like cell binders through a number of affinity selections, using whole cell protein extracts and membrane protein-enriched extracts from eight different GBM patients, and membrane protein-enriched extracts from two established GBM stem-like cell lines (NCH644 and NCH421K cells). After the enrichment, periplasmic extract ELISA was used to screen for specific clones. These nanobody clones were recloned into the pHEN6 vector, expressed in Escherichia coli WK6, and purified using immobilised metal affinity chromatography and size-exclusion chromatography. Specific nanobody:antigen pairs were obtained and mass spectrometry analysis revealed two proteins, TRIM28 and β-actin, that were up-regulated in the GBM stem-like cells compared to the controls.
Ključne besede:malignant gliomas, cancer stem cells, nanobodies
Status publikacije:Objavljeno
Verzija publikacije:Objavljena publikacija
Datum objave:24.11.2014
Leto izida:2014
Številčenje:Vol. 9, iss. 11
PID:20.500.12556/DiRROS-20012 Novo okno
UDK:620.3
ISSN pri članku:1932-6203
DOI:10.1371/journal.pone.0113688 Novo okno
COBISS.SI-ID:3251791 Novo okno
Opomba:Nasl. z nasl. zaslona; Opis vira z dne 25. 11. 2014; Članek v pdf obsega 22 str.;
Datum objave v DiRROS:02.08.2024
Število ogledov:438
Število prenosov:290
Metapodatki:XML DC-XML DC-RDF
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Gradivo je del revije

Naslov:PloS one
Založnik:Public Library of Science
ISSN:1932-6203
COBISS.SI-ID:2005896 Novo okno

Gradivo je financirano iz projekta

Financer:ARIS - Javna agencija za znanstvenoraziskovalno in inovacijsko dejavnost Republike Slovenije
Številka projekta:P1-0104-2009
Naslov:Funkcijska genomika in biotehnologija za zdravje

Financer:EC - European Commission
Številka projekta:205819
Naslov:Molecular Mechanisms of Glioma Genesis and Progression
Akronim:GLIOMA

Financer:Drugi - Drug financer ali več financerjev
Program financ.:INTERREG EC Project 2011
Številka projekta:42
Naslov:Identification of novel biomarkers to brain
Akronim:GLIOMA

Financer:Drugi - Drug financer ali več financerjev
Številka projekta:ASTF 26-2013
Naslov:European Molecular Biology Organisation
Akronim:EMBO

Financer:Drugi - Drug financer ali več financerjev
Program financ.:Slovene Human Resources Development and Scholarship Fund
Številka projekta:113

Licence

Licenca:CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
Povezava:http://creativecommons.org/licenses/by/4.0/deed.sl
Opis:To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

Jezik:Slovenski jezik
Ključne besede:maligni gliomi, izvorne rakave celice, nano telesa


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