Biotechnology and Bioprocess Engineering 2024; 29(3): 505-512  
Extraction of the outer membrane protein pertactin from Bordetella pertussis with urea for the production of acellular pertussis vaccine
Jae Hoon Moon1,2 · Jong Kwan Park2 · Bu Young Park2 · Hyung Jin Jeon2 · Gi Sub Choi2 · Gyun Min Lee1
1 Department of Biological Sciences, KAIST, Daejeon 34141, Korea
2 GC Biopharma, Yongin 16924, Korea
Correspondence to: Jae Hoon Moon
Moonandsky78@gmail.com
Gyun Min Lee
gyunminlee@kaist.ac.kr
Received: November 23, 2023; Revised: January 29, 2024; Accepted: February 6, 2024; Published online: June 10, 2024.
© The Korean Society for Biotechnology and Bioengineering. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
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.
Keywords: Pertactin · Acellular pertussis vaccine · Outer membrane protein · Urea extraction · Membrane protein purification

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