Human Monoclonal scFvs that Neutralize Fribrinogenolytic Activity of Kaouthiagin, a Zinc-Metalloproteinase in Cobra (<i>Naja kaouthia</i>) Venom
Snake venom-metalloproteinases (SVMPs) are the primary factors that disturb hemostasis and cause hemorrhage in the venomous snake bitten subjects. Kaouthiagin is a unique SVMP that binds and cleaves von Willebrand factor (vWF) at a specific peptide bond leading to inhibition of platelet aggregation,...
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2018-12-01
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doaj-156ab22d75554cc38fc1a707e9d102b62020-11-24T21:22:12ZengMDPI AGToxins2072-66512018-12-01101250910.3390/toxins10120509toxins10120509Human Monoclonal scFvs that Neutralize Fribrinogenolytic Activity of Kaouthiagin, a Zinc-Metalloproteinase in Cobra (<i>Naja kaouthia</i>) VenomJirawat Khanongnoi0Siratcha Phanthong1Onrapak Reamtong2Anchalee Tungtronchitr3Wanpen Chaicumpa4Nitat Sookrung5Graduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandGraduate Program in Immunology, Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandDepartment of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, ThailandCenter of Research Excellence on Therapeutic Proteins and Antibody Engineering, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandCenter of Research Excellence on Therapeutic Proteins and Antibody Engineering, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandCenter of Research Excellence on Therapeutic Proteins and Antibody Engineering, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, ThailandSnake venom-metalloproteinases (SVMPs) are the primary factors that disturb hemostasis and cause hemorrhage in the venomous snake bitten subjects. Kaouthiagin is a unique SVMP that binds and cleaves von Willebrand factor (vWF) at a specific peptide bond leading to inhibition of platelet aggregation, which enhances the hemorrhage. Kaouthiagin is a low abundant venom component of Thai cobra (<i>Naja kaouthia</i>); thus, most horse-derived antivenins used for cobra bite treatment do not contain adequate anti-kaouthiagin. This study aimed to produce human single-chain antibody variable fragments (HuscFvs) that bind to and interfere with kaouthiagin activity for further clinical use. Kaouthiagin was purified from <i>N. kaouthia</i>-holovenom by a single-step gel-filtration chromatography. The purified venom component was used in phage-biopanning to select the kaouthiagin-bound HuscFv-displayed-phage clones from a HuscFv-phage display library. The selected phages were used to infect Escherichia coli bacteria. Soluble HuscFvs expressed by three phage-transformed-<i>E. coli</i> clones interfered with cobra kaouthiagin binding to human vWF. Computerized simulation indicated that HuscFv of two phage-transformed <i>E. coli</i> clones formed contact interface with kaouthiagin residues at or near catalytic site and effectively inhibited fibrinogenolytic activity of the kaouthiagin. The HuscFvs have therapeutic potential as an adjunct of antivenins in treatment of bleeding caused by venomous snakebites.https://www.mdpi.com/2072-6651/10/12/509cobrahuman single-chain antibody variable fragments (HuscFvs)kaouthiagin<i>Naja kaouthia</i>snake venom metalloproteinase (SVMP)von Willebrand factor (vWF) |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jirawat Khanongnoi Siratcha Phanthong Onrapak Reamtong Anchalee Tungtronchitr Wanpen Chaicumpa Nitat Sookrung |
spellingShingle |
Jirawat Khanongnoi Siratcha Phanthong Onrapak Reamtong Anchalee Tungtronchitr Wanpen Chaicumpa Nitat Sookrung Human Monoclonal scFvs that Neutralize Fribrinogenolytic Activity of Kaouthiagin, a Zinc-Metalloproteinase in Cobra (<i>Naja kaouthia</i>) Venom Toxins cobra human single-chain antibody variable fragments (HuscFvs) kaouthiagin <i>Naja kaouthia</i> snake venom metalloproteinase (SVMP) von Willebrand factor (vWF) |
author_facet |
Jirawat Khanongnoi Siratcha Phanthong Onrapak Reamtong Anchalee Tungtronchitr Wanpen Chaicumpa Nitat Sookrung |
author_sort |
Jirawat Khanongnoi |
title |
Human Monoclonal scFvs that Neutralize Fribrinogenolytic Activity of Kaouthiagin, a Zinc-Metalloproteinase in Cobra (<i>Naja kaouthia</i>) Venom |
title_short |
Human Monoclonal scFvs that Neutralize Fribrinogenolytic Activity of Kaouthiagin, a Zinc-Metalloproteinase in Cobra (<i>Naja kaouthia</i>) Venom |
title_full |
Human Monoclonal scFvs that Neutralize Fribrinogenolytic Activity of Kaouthiagin, a Zinc-Metalloproteinase in Cobra (<i>Naja kaouthia</i>) Venom |
title_fullStr |
Human Monoclonal scFvs that Neutralize Fribrinogenolytic Activity of Kaouthiagin, a Zinc-Metalloproteinase in Cobra (<i>Naja kaouthia</i>) Venom |
title_full_unstemmed |
Human Monoclonal scFvs that Neutralize Fribrinogenolytic Activity of Kaouthiagin, a Zinc-Metalloproteinase in Cobra (<i>Naja kaouthia</i>) Venom |
title_sort |
human monoclonal scfvs that neutralize fribrinogenolytic activity of kaouthiagin, a zinc-metalloproteinase in cobra (<i>naja kaouthia</i>) venom |
publisher |
MDPI AG |
series |
Toxins |
issn |
2072-6651 |
publishDate |
2018-12-01 |
description |
Snake venom-metalloproteinases (SVMPs) are the primary factors that disturb hemostasis and cause hemorrhage in the venomous snake bitten subjects. Kaouthiagin is a unique SVMP that binds and cleaves von Willebrand factor (vWF) at a specific peptide bond leading to inhibition of platelet aggregation, which enhances the hemorrhage. Kaouthiagin is a low abundant venom component of Thai cobra (<i>Naja kaouthia</i>); thus, most horse-derived antivenins used for cobra bite treatment do not contain adequate anti-kaouthiagin. This study aimed to produce human single-chain antibody variable fragments (HuscFvs) that bind to and interfere with kaouthiagin activity for further clinical use. Kaouthiagin was purified from <i>N. kaouthia</i>-holovenom by a single-step gel-filtration chromatography. The purified venom component was used in phage-biopanning to select the kaouthiagin-bound HuscFv-displayed-phage clones from a HuscFv-phage display library. The selected phages were used to infect Escherichia coli bacteria. Soluble HuscFvs expressed by three phage-transformed-<i>E. coli</i> clones interfered with cobra kaouthiagin binding to human vWF. Computerized simulation indicated that HuscFv of two phage-transformed <i>E. coli</i> clones formed contact interface with kaouthiagin residues at or near catalytic site and effectively inhibited fibrinogenolytic activity of the kaouthiagin. The HuscFvs have therapeutic potential as an adjunct of antivenins in treatment of bleeding caused by venomous snakebites. |
topic |
cobra human single-chain antibody variable fragments (HuscFvs) kaouthiagin <i>Naja kaouthia</i> snake venom metalloproteinase (SVMP) von Willebrand factor (vWF) |
url |
https://www.mdpi.com/2072-6651/10/12/509 |
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