Production of Human Pancreatitis-Associated Protein (hPAP) in Komagataella phaffii (Pichia pastoris)
Pancreatitis-associated protein (PAP) is a pancreatic stress protein that is not produced in a healthy pancreas but is highly synthesized in pancreatic acinar cells in response to acute and chronic pancreatitis, hypoxia, toxins, diabetes, lipopolysaccharides hypotransferrinemia and organ transp...
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| Format: | Article |
| Language: | English |
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University of Sarajevo, Institute for Genetic Engineering and Biotechnology
2021-12-01
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| Series: | Genetics & Applications |
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| Online Access: | https://genapp.ba/editions/index.php/journal/article/view/146 |
| _version_ | 1811311592390787072 |
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| author | Didem Pazarli Fatıma Yücel Esin Akçael Şerife Şeyda Pirinçci Göktürk |
| author_facet | Didem Pazarli Fatıma Yücel Esin Akçael Şerife Şeyda Pirinçci Göktürk |
| author_sort | Didem Pazarli |
| collection | DOAJ |
| description |
Pancreatitis-associated protein (PAP) is a pancreatic stress protein that is not produced in a healthy pancreas but is highly synthesized in pancreatic acinar cells in response to acute and chronic pancreatitis, hypoxia, toxins, diabetes, lipopolysaccharides hypotransferrinemia and organ transplantation. Changes in the PAP levels in serum are an important biological marker in the early stage of pancreatic diseases. In this study, the recombinant human PAP protein, which has the potential to be used as a diagnostic marker and as research material in proliferation, apoptosis, cell migration, cell invasion, and immunoassay studies, was expressed efficiently under the control of the AOX1 gene promoter in the Komagataella phaffii (Pichia pastoris) (K. phaffii) X33 strain. We describe the conditions required for the efficient production of PAP protein by methanol induction and its use without purification. The produced unpurified protein was tested in sandwich ELISA and showed consistent results with the commercial product. These results are encouraging that the protein produced can be used as a biomarker standard in ELISA tests without the cost and labor of purification.
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| first_indexed | 2024-04-13T10:21:35Z |
| format | Article |
| id | doaj.art-32808901a428476788b61d3dd05ff9cd |
| institution | Directory Open Access Journal |
| issn | 2566-2937 2566-431X |
| language | English |
| last_indexed | 2024-04-13T10:21:35Z |
| publishDate | 2021-12-01 |
| publisher | University of Sarajevo, Institute for Genetic Engineering and Biotechnology |
| record_format | Article |
| series | Genetics & Applications |
| spelling | doaj.art-32808901a428476788b61d3dd05ff9cd2022-12-22T02:50:29ZengUniversity of Sarajevo, Institute for Genetic Engineering and BiotechnologyGenetics & Applications2566-29372566-431X2021-12-015210.31383/ga.vol5iss2pp31-39Production of Human Pancreatitis-Associated Protein (hPAP) in Komagataella phaffii (Pichia pastoris)Didem Pazarli0Fatıma Yücel1Esin Akçael2Şerife Şeyda Pirinçci Göktürk3Department of Medical Genetics and Molecular Biology, Institute of Health Sciences, Faculty of Medicine, Kocaeli University, Kocaeli, Turkey; The Scientific and Technological Research Council of Turkey (TÜBİTAK), Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli, TurkeyThe Scientific and Technological Research Council of Turkey (TÜBİTAK), Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli, TurkeyThe Scientific and Technological Research Council of Turkey (TÜBİTAK), Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli, Turkey The Scientific and Technological Research Council of Turkey (TÜBİTAK), Marmara Research Center, Genetic Engineering and Biotechnology Institute, Gebze, Kocaeli, Turkey Pancreatitis-associated protein (PAP) is a pancreatic stress protein that is not produced in a healthy pancreas but is highly synthesized in pancreatic acinar cells in response to acute and chronic pancreatitis, hypoxia, toxins, diabetes, lipopolysaccharides hypotransferrinemia and organ transplantation. Changes in the PAP levels in serum are an important biological marker in the early stage of pancreatic diseases. In this study, the recombinant human PAP protein, which has the potential to be used as a diagnostic marker and as research material in proliferation, apoptosis, cell migration, cell invasion, and immunoassay studies, was expressed efficiently under the control of the AOX1 gene promoter in the Komagataella phaffii (Pichia pastoris) (K. phaffii) X33 strain. We describe the conditions required for the efficient production of PAP protein by methanol induction and its use without purification. The produced unpurified protein was tested in sandwich ELISA and showed consistent results with the commercial product. These results are encouraging that the protein produced can be used as a biomarker standard in ELISA tests without the cost and labor of purification. https://genapp.ba/editions/index.php/journal/article/view/146Recombinant DNA TechnologyPancreatitis Associated ProteinPAPPichia pastorisELISAFusion Protein |
| spellingShingle | Didem Pazarli Fatıma Yücel Esin Akçael Şerife Şeyda Pirinçci Göktürk Production of Human Pancreatitis-Associated Protein (hPAP) in Komagataella phaffii (Pichia pastoris) Genetics & Applications Recombinant DNA Technology Pancreatitis Associated Protein PAP Pichia pastoris ELISA Fusion Protein |
| title | Production of Human Pancreatitis-Associated Protein (hPAP) in Komagataella phaffii (Pichia pastoris) |
| title_full | Production of Human Pancreatitis-Associated Protein (hPAP) in Komagataella phaffii (Pichia pastoris) |
| title_fullStr | Production of Human Pancreatitis-Associated Protein (hPAP) in Komagataella phaffii (Pichia pastoris) |
| title_full_unstemmed | Production of Human Pancreatitis-Associated Protein (hPAP) in Komagataella phaffii (Pichia pastoris) |
| title_short | Production of Human Pancreatitis-Associated Protein (hPAP) in Komagataella phaffii (Pichia pastoris) |
| title_sort | production of human pancreatitis associated protein hpap in komagataella phaffii pichia pastoris |
| topic | Recombinant DNA Technology Pancreatitis Associated Protein PAP Pichia pastoris ELISA Fusion Protein |
| url | https://genapp.ba/editions/index.php/journal/article/view/146 |
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