Cloning, expression, purification and functional analysis of a specific multi-epitope protein from multi drug resistance Acinetobacter baumannii
Background and Objectives: Immunization is a promising strategy to combat against the life-threatening infections by Multi Drug Resistance Acinetobacter baumannii. In this study, we directed to design and evaluate the efficacy of a recombinant multi-epitope protein against this pathogen. Materials a...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Tehran University of Medical Sciences
2021-10-01
|
| Series: | Iranian Journal of Microbiology |
| Subjects: | |
| Online Access: | https://ijm.tums.ac.ir/index.php/ijm/article/view/3112 |
| _version_ | 1818976064151486464 |
|---|---|
| author | Zahra Davoudi Amirhossein Taromchi Bahram Kazemi Mojgan Bandehpour Nariman Mosaffa |
| author_facet | Zahra Davoudi Amirhossein Taromchi Bahram Kazemi Mojgan Bandehpour Nariman Mosaffa |
| author_sort | Zahra Davoudi |
| collection | DOAJ |
| description | Background and Objectives: Immunization is a promising strategy to combat against the life-threatening infections by Multi Drug Resistance Acinetobacter baumannii. In this study, we directed to design and evaluate the efficacy of a recombinant multi-epitope protein against this pathogen.
Materials and Methods: Epitopes prediction was performed for candidate proteins OmpA and BAM complex (BamA, BamB, BamC, BamD, BamE) from A. baumannii, using immune-informatics tools with high affinity for the human HLA alleles. After expression and purification of the recombinant protein, its functional activity was confirmed by interaction with positive sera.
Results: Cloning and expression of the desired multi-epitopes protein were verified. Circular Dichroism study showed the secondary structure and proper refolding of the recombinant protein was achieved and matched with computational prediction. There was a significant interaction between designed protein with antibodies presented in ICU patients' and staff's sera.
Conclusion: The interaction of the recombinant protein with patients' sera antibodies suggests that it may be a promising determinant protein for immunization against of MDR A. baumannii. |
| first_indexed | 2024-12-20T16:05:54Z |
| format | Article |
| id | doaj.art-322edd15449b4fd5a8c0532c2ab215f1 |
| institution | Directory Open Access Journal |
| issn | 2008-3289 2008-4447 |
| language | English |
| last_indexed | 2024-12-20T16:05:54Z |
| publishDate | 2021-10-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Iranian Journal of Microbiology |
| spelling | doaj.art-322edd15449b4fd5a8c0532c2ab215f12022-12-21T19:34:08ZengTehran University of Medical SciencesIranian Journal of Microbiology2008-32892008-44472021-10-0113510.18502/ijm.v13i5.7429Cloning, expression, purification and functional analysis of a specific multi-epitope protein from multi drug resistance Acinetobacter baumanniiZahra Davoudi0Amirhossein Taromchi1Bahram Kazemi2Mojgan Bandehpour3Nariman Mosaffa4Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, IranDepartment of Medical Biotechnology and Nanotechnology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, IranCellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IranDepartment of Medical Biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran; Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, IranDepartment of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, IranBackground and Objectives: Immunization is a promising strategy to combat against the life-threatening infections by Multi Drug Resistance Acinetobacter baumannii. In this study, we directed to design and evaluate the efficacy of a recombinant multi-epitope protein against this pathogen. Materials and Methods: Epitopes prediction was performed for candidate proteins OmpA and BAM complex (BamA, BamB, BamC, BamD, BamE) from A. baumannii, using immune-informatics tools with high affinity for the human HLA alleles. After expression and purification of the recombinant protein, its functional activity was confirmed by interaction with positive sera. Results: Cloning and expression of the desired multi-epitopes protein were verified. Circular Dichroism study showed the secondary structure and proper refolding of the recombinant protein was achieved and matched with computational prediction. There was a significant interaction between designed protein with antibodies presented in ICU patients' and staff's sera. Conclusion: The interaction of the recombinant protein with patients' sera antibodies suggests that it may be a promising determinant protein for immunization against of MDR A. baumannii.https://ijm.tums.ac.ir/index.php/ijm/article/view/3112Multi drug resistant;Acinetobacter baumannii;Recombinant multi-epitope protein (rMEP);OmpA;BAM complex |
| spellingShingle | Zahra Davoudi Amirhossein Taromchi Bahram Kazemi Mojgan Bandehpour Nariman Mosaffa Cloning, expression, purification and functional analysis of a specific multi-epitope protein from multi drug resistance Acinetobacter baumannii Iranian Journal of Microbiology Multi drug resistant; Acinetobacter baumannii; Recombinant multi-epitope protein (rMEP); OmpA; BAM complex |
| title | Cloning, expression, purification and functional analysis of a specific multi-epitope protein from multi drug resistance Acinetobacter baumannii |
| title_full | Cloning, expression, purification and functional analysis of a specific multi-epitope protein from multi drug resistance Acinetobacter baumannii |
| title_fullStr | Cloning, expression, purification and functional analysis of a specific multi-epitope protein from multi drug resistance Acinetobacter baumannii |
| title_full_unstemmed | Cloning, expression, purification and functional analysis of a specific multi-epitope protein from multi drug resistance Acinetobacter baumannii |
| title_short | Cloning, expression, purification and functional analysis of a specific multi-epitope protein from multi drug resistance Acinetobacter baumannii |
| title_sort | cloning expression purification and functional analysis of a specific multi epitope protein from multi drug resistance acinetobacter baumannii |
| topic | Multi drug resistant; Acinetobacter baumannii; Recombinant multi-epitope protein (rMEP); OmpA; BAM complex |
| url | https://ijm.tums.ac.ir/index.php/ijm/article/view/3112 |
| work_keys_str_mv | AT zahradavoudi cloningexpressionpurificationandfunctionalanalysisofaspecificmultiepitopeproteinfrommultidrugresistanceacinetobacterbaumannii AT amirhosseintaromchi cloningexpressionpurificationandfunctionalanalysisofaspecificmultiepitopeproteinfrommultidrugresistanceacinetobacterbaumannii AT bahramkazemi cloningexpressionpurificationandfunctionalanalysisofaspecificmultiepitopeproteinfrommultidrugresistanceacinetobacterbaumannii AT mojganbandehpour cloningexpressionpurificationandfunctionalanalysisofaspecificmultiepitopeproteinfrommultidrugresistanceacinetobacterbaumannii AT narimanmosaffa cloningexpressionpurificationandfunctionalanalysisofaspecificmultiepitopeproteinfrommultidrugresistanceacinetobacterbaumannii |