Extraction of the outer membrane protein pertactin from Bordetella pertussis with urea for the production of acellular pertussis vaccine

• Published in: Biotechnology and bioprocess engineering Vol. 29; no. 3; pp. 505 - 512
• Main Authors: Moon, Jae Hoon, Park, Jong Kwan, Park, Bu Young, Jeon, Hyung Jin, Choi, Gi Sub, Lee, Gyun Min
• Format: Journal Article
• Language: English
• Published: Seoul The Korean Society for Biotechnology and Bioengineering 01.06.2024; Springer Nature B.V; 한국생물공학회
• Subjects: Anion exchange; Anion exchanging; Biotechnology; Bordetella pertussis; Cell culture; Chemistry; Chemistry and Materials Science; Freeze-thaw; Freezing; Gel chromatography; Gel filtration; Hydrophobicity; Impurities; Industrial and Production Engineering; Melting; Membrane proteins; Membranes; Pellets; Pertussis; Proteins; Research Paper; Thawing; Urea; Ureas; Vaccines; Whooping cough; 생물공학; Membrane protein purification; Pertactin; Acellular pertussis vaccine; Outer membrane protein; Urea extraction
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-024-00028-2

Pertactin (PRN), a non-fimbrial outer membrane protein of Bordetella pertussis , is the limiting component of the acellular pertussis vaccine because of its low concentration. This study aimed to develop a large-scale urea-based process for PRN extraction from B. pertussis . Cell pellet processing conditions, including freezing and thawing, were found to substantially affect PRN yield. A single cycle of rapid freezing of the cell pellet at − 30 °C with slow thawing at 5 ± 3 °C resulted in up to fivefold higher PRN yield than condition without freezing and thawing. The search for urea treatment conditions was also conducted, and 5 M urea treatment for 2 h was the optimal condition. The developed urea-based process was applied to 50 L culture scale, and residual impurities were removed by sequential anion exchange, hydrophobic interaction and gel filtration chromatography and resulted in PRN with a purity of over 95% at a yield of 33.2%. From 50 L culture broth, the final yield of PRN per cell pellet was 0.23 mg/g (wet weight). Thus, a large-scale production process for high-quality PRN from B. pertussis was developed based on urea extraction process. The results may serve as a reference for production of other membrane proteins.