Novel mouse monoclonal antibodies against Bordetella pertussis pertactin antigen with versatile applications
Copyright © 2023 Elsevier B.V. All rights reserved..
BACKGROUND: Pertussis, or whooping cough, is a highly contagious respiratory disease caused by Bordetella pertussis (BP). Pertactin (PRN) is one of the main immunogenic components of BP and is employed in many commercialized acellular pertussis vaccines (aPVs). Purification of this protein by conventional chromatography methods is challenging and commonly requires multiple laborious processes with low recovery. Using specific monoclonal antibodies (mAbs) for the purification of PRN antigen is expected to yield high purity and recovery of the target molecule.
METHODS: Recombinant PRN antigen was used to produce mouse mAbs using hybridoma technology. Structural and functional characteristics of the mAbs were assessed by ELISA, immunoblotting, and flow cytometry. Selected mAbs were employed to purify PRN by affinity chromatography, and the purity and recovery of the purified protein were analyzed by ELISA, SDS-PAGE, and immunoblotting. Moreover, ELISA and flow cytometry techniques were designed using these mAbs to detect PRN in different strains of BP.
RESULTS: Five mAbs were produced and selected based on their reactivity with native PRN. Our results demonstrate that purification of PRN by affinity chromatography resulted in a highly pure antigen with 75-85 percent recovery. In addition, ELISA and flow cytometry results indicated that these mAbs could recognize PRN in the bacterial cell walls of different BP strains.
CONCLUSION: We successfully produced PRN-specific mAbs and designed an affinity chromatography method to purify PRN antigen with higher purity and recovery than conventional methods. These mAbs could be employed as valuable tools for the detection and purification of PRN for vaccine manufacturing.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:211 |
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| Enthalten in: |
Journal of microbiological methods - 211(2023) vom: 24. Aug., Seite 106786 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Imani, Danyal [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 08.08.2023 Date Revised 08.08.2023 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1016/j.mimet.2023.106786 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| 245 | 1 | 0 | |a Novel mouse monoclonal antibodies against Bordetella pertussis pertactin antigen with versatile applications |
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| 500 | |a Date Revised 08.08.2023 | ||
| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2023 Elsevier B.V. All rights reserved. | ||
| 520 | |a BACKGROUND: Pertussis, or whooping cough, is a highly contagious respiratory disease caused by Bordetella pertussis (BP). Pertactin (PRN) is one of the main immunogenic components of BP and is employed in many commercialized acellular pertussis vaccines (aPVs). Purification of this protein by conventional chromatography methods is challenging and commonly requires multiple laborious processes with low recovery. Using specific monoclonal antibodies (mAbs) for the purification of PRN antigen is expected to yield high purity and recovery of the target molecule | ||
| 520 | |a METHODS: Recombinant PRN antigen was used to produce mouse mAbs using hybridoma technology. Structural and functional characteristics of the mAbs were assessed by ELISA, immunoblotting, and flow cytometry. Selected mAbs were employed to purify PRN by affinity chromatography, and the purity and recovery of the purified protein were analyzed by ELISA, SDS-PAGE, and immunoblotting. Moreover, ELISA and flow cytometry techniques were designed using these mAbs to detect PRN in different strains of BP | ||
| 520 | |a RESULTS: Five mAbs were produced and selected based on their reactivity with native PRN. Our results demonstrate that purification of PRN by affinity chromatography resulted in a highly pure antigen with 75-85 percent recovery. In addition, ELISA and flow cytometry results indicated that these mAbs could recognize PRN in the bacterial cell walls of different BP strains | ||
| 520 | |a CONCLUSION: We successfully produced PRN-specific mAbs and designed an affinity chromatography method to purify PRN antigen with higher purity and recovery than conventional methods. These mAbs could be employed as valuable tools for the detection and purification of PRN for vaccine manufacturing | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a Affinity chromatography | |
| 650 | 4 | |a Bordetella pertussis | |
| 650 | 4 | |a Monoclonal antibody | |
| 650 | 4 | |a Pertactin | |
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| 650 | 7 | |a Virulence Factors, Bordetella |2 NLM | |
| 650 | 7 | |a Bacterial Outer Membrane Proteins |2 NLM | |
| 650 | 7 | |a Pertussis Vaccine |2 NLM | |
| 650 | 7 | |a Antibodies, Monoclonal |2 NLM | |
| 650 | 7 | |a Antibodies, Bacterial |2 NLM | |
| 700 | 1 | |a Bahadori, Tannaz |e verfasserin |4 aut | |
| 700 | 1 | |a Ghourchian, Sedighe |e verfasserin |4 aut | |
| 700 | 1 | |a Golsaz-Shirazi, Forough |e verfasserin |4 aut | |
| 700 | 1 | |a Douraghi, Masoumeh |e verfasserin |4 aut | |
| 700 | 1 | |a Jeddi-Tehrani, Mahmood |e verfasserin |4 aut | |
| 700 | 1 | |a Amiri, Mohammad Mehdi |e verfasserin |4 aut | |
| 700 | 1 | |a Shokri, Fazel |e verfasserin |4 aut | |
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