@article{MTMT:35434014, title = {Development of a Mammalian Cell Line for Stable Production of Anti-PD-1}, url = {https://m2.mtmt.hu/api/publication/35434014}, author = {Csató-Kovács, Erika and Salamon, Pál Attila and Fikó-Lászlo, Szilvia and Kovács, Krisztina and Koka, Alice and András-Korodi, Mónika and Antal, Emőke and Brumă, Emília and Tőrsők, Brigitta and Gudor, Szilárd and Miklóssy, Ildikó and Orbán, Kálmán Csongor and Albert, Csilla and Bálint, Emese Éva and Albert, Beáta}, doi = {10.3390/antib13040082}, journal-iso = {ANTIBODIES}, journal = {ANTIBODIES}, volume = {13}, unique-id = {35434014}, abstract = {Background/Objectives: Immune checkpoint blockade, particularly targeting the programmed cell death 1 (PD-1) receptor, is a promising strategy in cancer immunotherapy. The interaction between PD-1 and its ligands, PD-L1 and PD-L2, is crucial in immune evasion by tumors. Blocking this interaction with monoclonal antibodies like Nivolumab can restore anti-tumor immunity. This study aims to develop a stable expression system for Nivolumab-based anti-PD-1 in the Chinese Hamster Ovary (CHO) DG44 cell line using two different expression vector systems with various signal sequences. Methods: The heavy chain (HC) and light chain (LC) of Nivolumab were cloned into two expression vectors, pOptiVEC and pcDNA3.3. Each vector was engineered with two distinct signal sequences, resulting in the creation of eight recombinant plasmids. These plasmids were co-transfected into CHO DG44 cells in different combinations, allowing for the assessment of stable antibody production. Results: Both pOptiVEC and pcDNA3.3 vectors were successful in stably integrating and expressing the Nivolumab-based anti-PD-1 antibody in CHO DG44 cells. This study found that the choice of signal sequence significantly influenced the quantity of antibodies produced. The optimization of production conditions further enhanced antibody yield, indicating the potential for large-scale production. Conclusions: This study demonstrates that both pOptiVEC and pcDNA3.3 expression systems are effective for the stable production of Nivolumab-based anti-PD-1 in CHO DG44 cells. Signal sequences play a critical role in determining the expression levels, and optimizing production conditions can further increase antibody yield, supporting future applications in cancer immunotherapy.}, year = {2024}, eissn = {2073-4468}, orcid-numbers = {Salamon, Pál Attila/0000-0002-6803-1212; Miklóssy, Ildikó/0000-0002-8037-7216; Orbán, Kálmán Csongor/0000-0002-7509-748X; Albert, Csilla/0000-0001-6882-6691} } @article{MTMT:34238005, title = {Enhanced Gait Recovery in Chronic Post-COVID-19 Stroke: The Role of Combined Physical Rehabilitation}, url = {https://m2.mtmt.hu/api/publication/34238005}, author = {Fodor, Hunor Pál and Dávid, Hunor and Czont, Attila and Miklóssy, Ildikó and Orbán, Kálmán-Csongor and Tar, Gyöngyi and Fodor, Abony and Kovács, Zita and Albert, Beáta and Salamon, Pál Attila}, doi = {10.3390/reports6040051}, journal-iso = {REPORTS-BASEL}, journal = {REPORTS}, volume = {6}, unique-id = {34238005}, abstract = {Background: Rehabilitation programs applied in cases of COVID-19-related stroke should counteract not only the effects of the stroke but also the effects of long-term COVID-19. As the molecular processes underlying these cases are still not fully understood, and evidence-based clinical outcomes are scarcely documented, there is a valid need to gather information and develop rehabilitation strategies for these patients. The risks, already clarified in the case of stroke, need to be assessed taking into account the coincidence of the two diseases. Endothelial injuries and emboli that develop after the hypercoagulable state of COVID-19 may take longer to heal, and complications may occur during exercise. This case study attempts to determine what the rehabilitation of a COVID-19-related stroke patient should include. The participant was a 64-year-old male with ischemic right middle cerebral artery stroke, left-side hemiplegia, and middle cerebral artery stenosis, and the CT showed a well-defined area of hypoattenuation in the basal ganglia territory involving the right lentiform nucleus, the anterior and posterior limbs of the internal capsule, and the dorsal part of the external capsule. His NIHSS score was 14, and he registered 15 points on the Barthel index. The patient had a COVID-19 infection two weeks before the stroke event. Methods: Conventional physical therapy was combined with adaptive ballistic strength training, a high-intensity interval training regimen, and manual treatment for myofascial release throughout the chronic recovery phase. Our primary goals were gait rehabilitation, muscle strengthening, weakness management, as well as spasticity reduction, while three different rehabilitation approaches were adopted in a single rehabilitation program to improve the outcome and long-term functional recovery of the patient. Results: The patient progressed in almost every aspect of the assessment criteria. This combined approach’s main success was improved gait speed, gait quality, and improved cardiovascular fitness. Take-away message: In the case of a stroke caused by COVID-19, where the endothelium cells are compromised, HIIT may be questionable due to the poor vascular condition. Based on our results, the low-volume HIIT approach proved appropriate and effective.}, year = {2023}, eissn = {2571-841X}, orcid-numbers = {Dávid, Hunor/0009-0004-3834-421X; Czont, Attila/0000-0002-6395-1562; Miklóssy, Ildikó/0000-0002-8037-7216; Orbán, Kálmán-Csongor/0000-0002-7509-748X} } @article{MTMT:34109004, title = {THE EFFECT OF SARS-COV-2 VIRUS ON THE ACTIVITY OF THE PANOPTOSIS PATHWAY IN UPPER RESPIRATORY EPITHELIAL CELLS: CORRELATIONS BETWEEN EXPRESSION PATTERNS, AGE, GENDER, AND COMORBIDITIES}, url = {https://m2.mtmt.hu/api/publication/34109004}, author = {Kovács, Zita and Bálint, Emese-Éva and Lászlo, Szilvia and Albert, Beáta and Gyöngyi, Tar and Attila, András and Salamon, Pál Attila}, journal-iso = {ACTA MARISIEN}, journal = {ACTA MARISIENSIS: SERIA MEDICA}, volume = {69}, unique-id = {34109004}, issn = {2668-7755}, year = {2023}, eissn = {2668-7763}, pages = {35-35} } @article{MTMT:33365403, title = {Structural hierarchy of mechanical extensibility in human von Willebrand factor multimers}, url = {https://m2.mtmt.hu/api/publication/33365403}, author = {Csányi, Mária Csilla and Salamon, Pál Attila and Feller, Tímea and Bozó, Tamás and Hársfalvi, Jolán and Kellermayer, Miklós}, doi = {10.1002/pro.4535}, journal-iso = {PROTEIN SCI}, journal = {PROTEIN SCIENCE}, volume = {32}, unique-id = {33365403}, issn = {0961-8368}, year = {2023}, eissn = {1469-896X}, orcid-numbers = {Csányi, Mária Csilla/0000-0002-7777-4229; Bozó, Tamás/0000-0002-2643-0661; Hársfalvi, Jolán/0000-0001-9940-4846; Kellermayer, Miklós/0000-0002-5553-6553} } @CONFERENCE{MTMT:33550801, title = {Force dependent multimer glycoprotein elongation}, url = {https://m2.mtmt.hu/api/publication/33550801}, author = {Csányi, Mária Csilla and Salamon, Pál Attila and Feller, Tímea and Bozó, Tamás and Hársfalvi, Jolán and Kellermayer, Miklós S.Z.}, booktitle = {20th INTERNATIONAL SYMPOSIUM AND SUMMER SCHOOL ON BIOANALYSIS Book of Abstracts}, unique-id = {33550801}, year = {2022}, pages = {64-64}, orcid-numbers = {Csányi, Mária Csilla/0000-0002-7777-4229} } @article{MTMT:33345866, title = {Study of native SMAC protein production in the pUbiq expression system: Molecular cloning, biosynthesis and molecular modelling}, url = {https://m2.mtmt.hu/api/publication/33345866}, author = {Salamon, Pál Attila and Orbán, Kálmán Csongor and Molnar-Nagy, Katalin and Kovács, Zita and Vancsa, Klara and Bálint, Emese-Éva and Miklossy, Ildiko and Albert, Beáta and Tar, Gyongyi and Lanyi, Szabolcs}, doi = {10.1016/j.ejbt.2022.01.0020717-3458}, journal-iso = {ELECTRON J BIOTECHN}, journal = {ELECTRONIC JOURNAL OF BIOTECHNOLOGY}, volume = {56}, unique-id = {33345866}, issn = {0717-3458}, abstract = {Background: In the process of recombinant protein biosynthesis affinity tags are efficient tools to achieve the expected purity and yield during the purification steps. Nonetheless these tags might alter enzyme specificity and activity, therefore in functional assays it is recommended to use authentic or native proteins. Several ubiquitin fusion systems have been developed for E. coli-based recombinant protein expression that provide high levels of expression, with simple purification, and allow the production of various proteins with authentic N-terminus for subsequent applications. Results: In the present research, we describe an ubiquitin fused bacterial biosynthetic system (pUbiq) for the production of the native Second mitochondria-derived activator of caspases (SMAC) recombinant protein. Using this system, the recombinant protein is expressed with an ubiquitin-decahistidine fusion partner, then purified from the cell-forming proteins by affinity chromatography. The fusion partner is then removed by proteolytic digestion, resulting the native structure of the recombinant protein without unnecessary amino acid residues. Following proteolysis, another affinity chromatography method is used to separate the native protein from the fusion partner and the proteolytic enzyme. The folding of the protein of interest was verified by a pull-down assay. Conclusions: Based on our results, the presented pUbiq system was successfully applied in the production of native SMAC recombinant protein, where the affinity tag required for purification was completely removed. Our study suggests that the ubiquitin-fusion technology will be useful for enhancing expression and purification of native and authentic proteins for structural and functional studies as well as for therapeutic uses. How to cite: Salamona P, Orbana CK, Molnar-Nagy K, et al. Exon based amplified polymorphism (EBAP): A novel and universal molecular marker for plants. Electron J Biotechnol 2022;56. https://doi.org/10.10 16/j.ejbt.2022.01.002. (c) 2022 Universidad Catolica de Valparaiso. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).}, keywords = {SYSTEM; Molecular cloning; biosynthesis; recombinant protein; molecular modelling; caspases; affinity tags; Smac; Expression construct; Second mitochondria-derived activator of; Ubiquitin fused bacterial biosynthetic; Ubiquitin fusion systems}, year = {2022}, eissn = {0717-3458}, pages = {39-46} } @article{MTMT:32915059, title = {Quantitative and Qualitative Assessment of Fluorescence in Aesthetic Direct Restorations}, url = {https://m2.mtmt.hu/api/publication/32915059}, author = {Bardocz-Veres, Zsuzsanna and Székely, Melinda and Salamon, Pál Attila and Bala, Előd and Bereczki, Előd and Kerekes-Máthé, Bernadette}, doi = {10.3390/ma15134619}, journal-iso = {MATERIALS}, journal = {MATERIALS}, volume = {15}, unique-id = {32915059}, year = {2022}, eissn = {1996-1944}, pages = {4619}, orcid-numbers = {Székely, Melinda/0000-0002-3998-0638; Kerekes-Máthé, Bernadette/0000-0001-9643-6383} } @article{MTMT:32923364, title = {COMPARATIVE STUDY ON CONVENTIONAL AND AUTOINDUCTION FERMENTATION}, url = {https://m2.mtmt.hu/api/publication/32923364}, author = {Salamon, Pál Attila and Orbán, Kálmán Csongor and Ildikó, MIKLÓSSY and Albert, Beáta and Szabolcs, LÁNYI}, journal-iso = {UPB SCI BULL SER B}, journal = {UPB SCIENTIFIC BULLETIN, SERIES B: CHEMISTRY AND MATERIALS SCIENCE}, volume = {83}, unique-id = {32923364}, issn = {1454-2331}, year = {2021}, eissn = {1454-2331}, pages = {77} } @article{MTMT:32547005, title = {Detergent aided refolding and purification of recombinant XIAP from inclusion bodies}, url = {https://m2.mtmt.hu/api/publication/32547005}, author = {Nagy, Katalin and Kovács, Zita and Miklóssy, Ildikó and Salamon, Pál Attila and Orbán, Kálmán Csongor and Albert, Beáta and Lányi, Szabolcs}, doi = {10.24193/subbchem.2021.4.26}, journal-iso = {STUD UNIV BABES-BOLYAI CHEM}, journal = {STUDIA UNIVERSITATIS BABES-BOLYAI CHEMIA}, volume = {66}, unique-id = {32547005}, issn = {1224-7154}, year = {2021}, eissn = {2065-9520}, pages = {355-368} } @article{MTMT:32069722, title = {IN VITRO STUDY OF THE CCMV CAPSID PROTEIN: CLONING, EXPRESSION, AND PURIFICATION}, url = {https://m2.mtmt.hu/api/publication/32069722}, author = {János, SZÖVÉRFI and Orbán, Kálmán Csongor and Albert, Beáta and Katalin, NAGY and Salamon, Pál Attila and Szabolcs, LÁNYI}, journal-iso = {UPB SCI BULL SER B}, journal = {UPB SCIENTIFIC BULLETIN, SERIES B: CHEMISTRY AND MATERIALS SCIENCE}, volume = {83}, unique-id = {32069722}, issn = {1454-2331}, abstract = {In our current work we studied the expression of the Cowpea chlorotic mottle virus (CCMV) capsid protein in E. coli host cell. The CCMV proteins DNA was cloned into a vector containing a His-tag and a ubiquitin fusion partner. The gene was transformed in E. coli BL21(DE3) Rosetta cells and expressed. The expression process was optimized and shake flask expression was conducted on 37 degrees C and 0.1 mM IPTG concentration. Cloning and expressing of CCMV capsid protein was successful, confirmed by sequencing results. The protein was purified with Ni-affinity chromatography.}, year = {2021}, eissn = {1454-2331}, pages = {135-142} }