Single chain antibody fragment with serine protease inhibitory property capable of neutralizing toxicity of Trimeresurus mucrosquamatus venom

Yu Ching Lee, Wang Chuan Chen, Meng Huei Liang, Chi Hsin Lee, Keng Chang Tsai, Jen Ron Chiang, Liao Chun Chiang, Chi Ching Chen, Chang Yu Chang, Ching Hsiao Lee, Sy Jye Leu, Yi Yuan Yang

研究成果: 雜誌貢獻文章

2 引文 (Scopus)

摘要

Trimeresurus mucrosquamatus (TM) is one of majorities of snake envenomation with necrotic and hemorrhagic toxin in Taiwan. In this study, chickens were used as an alternative animal model for immunization with TM venom. Using phage display technology to process four rounds of panning, selected single chain variable fragments (scFv) could specifically recognize TM venom proteins, which were later identified as a group of homogeneous venom serine protease. The specific scFv antibodies showed various inhibitory effects on sheep RBC lysis induced by TM venom using an indirect hemolytic assay in vitro. In addition, the survival times of mice were extended to certain degrees when treated with these scFv antibodies individually or in a combination. To elucidate the inhibitory mechanism, we used molecular modeling to build up the serine protease structure to simulate the possible interactions with scFv antibodies. The results suggested that the CDR-loop of the scFv antibodies (3S10 or 4S1) might bind at the 99-loop of venom serine protease so as to affect substrate access due to the partial collapse of the subsite S2 and the partial movement of the subsite S4. It is hoped these chicken-derived antibodies could be applied to develop diagnostic and therapeutic agents against snakebites.

原文英語
頁(從 - 到)170-176
頁數7
期刊Biochemical and Biophysical Research Communications
460
發行號2
DOIs
出版狀態已發佈 - 五月 1 2015

指紋

Single-Chain Antibodies
Immunoglobulin Fragments
Serine Proteases
Toxicity
Snake Bites
Venoms
Chickens
Trimeresurus
Immunization
Bacteriophages
Molecular modeling
Taiwan
Assays
Sheep
Animals
Animal Models
Display devices
Trimeresurus venoms
Technology
Antibodies

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

引用此文

Single chain antibody fragment with serine protease inhibitory property capable of neutralizing toxicity of Trimeresurus mucrosquamatus venom. / Lee, Yu Ching; Chen, Wang Chuan; Liang, Meng Huei; Lee, Chi Hsin; Tsai, Keng Chang; Chiang, Jen Ron; Chiang, Liao Chun; Chen, Chi Ching; Chang, Chang Yu; Lee, Ching Hsiao; Leu, Sy Jye; Yang, Yi Yuan.

於: Biochemical and Biophysical Research Communications, 卷 460, 編號 2, 01.05.2015, p. 170-176.

研究成果: 雜誌貢獻文章

Lee, Yu Ching ; Chen, Wang Chuan ; Liang, Meng Huei ; Lee, Chi Hsin ; Tsai, Keng Chang ; Chiang, Jen Ron ; Chiang, Liao Chun ; Chen, Chi Ching ; Chang, Chang Yu ; Lee, Ching Hsiao ; Leu, Sy Jye ; Yang, Yi Yuan. / Single chain antibody fragment with serine protease inhibitory property capable of neutralizing toxicity of Trimeresurus mucrosquamatus venom. 於: Biochemical and Biophysical Research Communications. 2015 ; 卷 460, 編號 2. 頁 170-176.
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abstract = "Trimeresurus mucrosquamatus (TM) is one of majorities of snake envenomation with necrotic and hemorrhagic toxin in Taiwan. In this study, chickens were used as an alternative animal model for immunization with TM venom. Using phage display technology to process four rounds of panning, selected single chain variable fragments (scFv) could specifically recognize TM venom proteins, which were later identified as a group of homogeneous venom serine protease. The specific scFv antibodies showed various inhibitory effects on sheep RBC lysis induced by TM venom using an indirect hemolytic assay in vitro. In addition, the survival times of mice were extended to certain degrees when treated with these scFv antibodies individually or in a combination. To elucidate the inhibitory mechanism, we used molecular modeling to build up the serine protease structure to simulate the possible interactions with scFv antibodies. The results suggested that the CDR-loop of the scFv antibodies (3S10 or 4S1) might bind at the 99-loop of venom serine protease so as to affect substrate access due to the partial collapse of the subsite S2 and the partial movement of the subsite S4. It is hoped these chicken-derived antibodies could be applied to develop diagnostic and therapeutic agents against snakebites.",
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AU - Tsai, Keng Chang

AU - Chiang, Jen Ron

AU - Chiang, Liao Chun

AU - Chen, Chi Ching

AU - Chang, Chang Yu

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AU - Yang, Yi Yuan

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AB - Trimeresurus mucrosquamatus (TM) is one of majorities of snake envenomation with necrotic and hemorrhagic toxin in Taiwan. In this study, chickens were used as an alternative animal model for immunization with TM venom. Using phage display technology to process four rounds of panning, selected single chain variable fragments (scFv) could specifically recognize TM venom proteins, which were later identified as a group of homogeneous venom serine protease. The specific scFv antibodies showed various inhibitory effects on sheep RBC lysis induced by TM venom using an indirect hemolytic assay in vitro. In addition, the survival times of mice were extended to certain degrees when treated with these scFv antibodies individually or in a combination. To elucidate the inhibitory mechanism, we used molecular modeling to build up the serine protease structure to simulate the possible interactions with scFv antibodies. The results suggested that the CDR-loop of the scFv antibodies (3S10 or 4S1) might bind at the 99-loop of venom serine protease so as to affect substrate access due to the partial collapse of the subsite S2 and the partial movement of the subsite S4. It is hoped these chicken-derived antibodies could be applied to develop diagnostic and therapeutic agents against snakebites.

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