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Development of Monoclonal Antibodies Specific to Galectin of Pine Wood Nematode, Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle and Their Utilization for Detection of Pine Wood Nematodes

소나무재선충[Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle]의 GaLectin에 대한 특이적인 단클론 항체 제작과 진단에의 활용

  • Kim, A-Young (Ilsong Institute of Life Science, Hallym University) ;
  • Kim, Young Ha (Ilsong Institute of Life Science, Hallym University) ;
  • Choi, Bo-Hye (Ilsong Institute of Life Science, Hallym University) ;
  • Nguyen, Trang (Ilsong Institute of Life Science, Hallym University) ;
  • Yoon, Kyungjae Andrew (Department of Agricultural Biology, Seoul National University) ;
  • Lee, Si Hyeock (Department of Agricultural Biology, Seoul National University) ;
  • Han, Hye-Rim (Laboratory of Pine Wilt Disease, Division of Forest Insect Pests & Disease, National Institute of Forest Science) ;
  • Koh, Young Ho (Ilsong Institute of Life Science, Hallym University)
  • 김아영 (한림대학교 일송생명과학연구소) ;
  • 김영하 (한림대학교 일송생명과학연구소) ;
  • 최보혜 (한림대학교 일송생명과학연구소) ;
  • 트랑뉴겐 (한림대학교 일송생명과학연구소) ;
  • 윤경재 (서울대학교 농생물학과) ;
  • 이시혁 (서울대학교 농생물학과) ;
  • 한혜림 (국립산림과학원) ;
  • 고영호 (한림대학교 일송생명과학연구소)
  • Received : 2017.10.24
  • Accepted : 2018.01.16
  • Published : 2018.03.01

Abstract

Currently, there is no available tool that rapidly diagnoses pine wood nematode (PWN)-infected pine trees in the field. In this study, we synthesized and purified PWN Galectin, which might be an antigen specific to PWN, using the Baculovirus expression system. We used PWN Galectin as an antigen for generating 1,464 fusion hybridoma cell lines secreting monoclonal antibodies (Mabs). Among them, we selected 62 fusion hybridoma cell lines showing high reactivity to PWN Galectin. We further selected 12 fusion hybridoma cell lines showing high reactivity to the standard PWN-infected pine tree phosphate buffered saline (PBS) extract. Additionally, two fusion hybridoma cell lines showing no or extremely low reactivity were used as controls. The selected fusion hybridoma cell lines were subjected to limiting dilutions for selecting and establishing Mab-secreting cell lines showing higher reactivity to the standard PWN-infected pine tree extract than to the standard normal pine tree PBS extract. Moreover, the selected fusion hybridoma cell lines were further selected based on their higher reactivity to PWN protein extracts than to three non-pathogenic nematode protein extracts. The Mab-secreting cell lines established in this study could be used to develop rapid diagnostic tools that can be used in the field or in laboratories for detecting PWN-infected pine trees or PWN.

Keywords

Pine wood nematode;Galectin;Infected pine tree;Non-pathogenic pine tree;Mab-secreting cell line

Acknowledgement

Grant : 소나무재선충 종 특이 항원 및 항체를 이용한 진단 기술 개발

Supported by : 산림과학원

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