A novel in vitro potency assay of antisera against Thai Naja kaouthia based on nicotinic acetylcholine receptor binding
Abstract Snake envenomation is an important medical problem. One of the hurdles in antivenom development is the in vivo assay of antivenom potency which is expensive, gives variable results and kills many animals. We report a novel in vitro assay involving the specific binding of the postsynaptic ne...
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Nature Portfolio
2017
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oai:doaj.org-article:c37c2be9b26744c9a2ac701ad38d5c982021-12-02T15:04:51ZA novel in vitro potency assay of antisera against Thai Naja kaouthia based on nicotinic acetylcholine receptor binding10.1038/s41598-017-08962-32045-2322https://doaj.org/article/c37c2be9b26744c9a2ac701ad38d5c982017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08962-3https://doaj.org/toc/2045-2322Abstract Snake envenomation is an important medical problem. One of the hurdles in antivenom development is the in vivo assay of antivenom potency which is expensive, gives variable results and kills many animals. We report a novel in vitro assay involving the specific binding of the postsynaptic neurotoxins (PSNTs) of elapid snakes with purified Torpedo californica nicotinic acetylcholine receptor (nAChR). The potency of an antivenom is determined by its antibody ability to bind and neutralize the PSNT, thus preventing it from binding to nAChR. The PSNT of Naja kaouthia (NK3) was immobilized on microtiter wells and nAChR was added to bind with it. The in vitro IC50 of N. kaouthia venom that inhibited 50% of nAChR binding to the immobilized NK3 was determined. Varying concentrations of antisera against N. kaouthia were separately pre-incubated with 5xIC50 of N. kaouthia venom. The remaining free NK3 were incubated with nAChR before adding to the NK3 coated plates. The in vitro and in vivo median effective ratio, ER50s of 12 batches of antisera showed correlation (R 2) of 0.9809 (p < 0.0001). This in vitro assay should be applicable to antisera against other elapid venoms and should reduce the use of live animals and accelerate development of life-saving antivenoms.Kavi RatanabanangkoonPavinee SimsiriwongKritsada PruksaphonKae Yi TanSukanya EursakunChoo Hock TanBunkuea ChantrathonkulWongsakorn WongwadhunyooSirida YoungchimNget Hong TanNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-8 (2017) |
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Medicine R Science Q Kavi Ratanabanangkoon Pavinee Simsiriwong Kritsada Pruksaphon Kae Yi Tan Sukanya Eursakun Choo Hock Tan Bunkuea Chantrathonkul Wongsakorn Wongwadhunyoo Sirida Youngchim Nget Hong Tan A novel in vitro potency assay of antisera against Thai Naja kaouthia based on nicotinic acetylcholine receptor binding |
| description |
Abstract Snake envenomation is an important medical problem. One of the hurdles in antivenom development is the in vivo assay of antivenom potency which is expensive, gives variable results and kills many animals. We report a novel in vitro assay involving the specific binding of the postsynaptic neurotoxins (PSNTs) of elapid snakes with purified Torpedo californica nicotinic acetylcholine receptor (nAChR). The potency of an antivenom is determined by its antibody ability to bind and neutralize the PSNT, thus preventing it from binding to nAChR. The PSNT of Naja kaouthia (NK3) was immobilized on microtiter wells and nAChR was added to bind with it. The in vitro IC50 of N. kaouthia venom that inhibited 50% of nAChR binding to the immobilized NK3 was determined. Varying concentrations of antisera against N. kaouthia were separately pre-incubated with 5xIC50 of N. kaouthia venom. The remaining free NK3 were incubated with nAChR before adding to the NK3 coated plates. The in vitro and in vivo median effective ratio, ER50s of 12 batches of antisera showed correlation (R 2) of 0.9809 (p < 0.0001). This in vitro assay should be applicable to antisera against other elapid venoms and should reduce the use of live animals and accelerate development of life-saving antivenoms. |
| format |
article |
| author |
Kavi Ratanabanangkoon Pavinee Simsiriwong Kritsada Pruksaphon Kae Yi Tan Sukanya Eursakun Choo Hock Tan Bunkuea Chantrathonkul Wongsakorn Wongwadhunyoo Sirida Youngchim Nget Hong Tan |
| author_facet |
Kavi Ratanabanangkoon Pavinee Simsiriwong Kritsada Pruksaphon Kae Yi Tan Sukanya Eursakun Choo Hock Tan Bunkuea Chantrathonkul Wongsakorn Wongwadhunyoo Sirida Youngchim Nget Hong Tan |
| author_sort |
Kavi Ratanabanangkoon |
| title |
A novel in vitro potency assay of antisera against Thai Naja kaouthia based on nicotinic acetylcholine receptor binding |
| title_short |
A novel in vitro potency assay of antisera against Thai Naja kaouthia based on nicotinic acetylcholine receptor binding |
| title_full |
A novel in vitro potency assay of antisera against Thai Naja kaouthia based on nicotinic acetylcholine receptor binding |
| title_fullStr |
A novel in vitro potency assay of antisera against Thai Naja kaouthia based on nicotinic acetylcholine receptor binding |
| title_full_unstemmed |
A novel in vitro potency assay of antisera against Thai Naja kaouthia based on nicotinic acetylcholine receptor binding |
| title_sort |
novel in vitro potency assay of antisera against thai naja kaouthia based on nicotinic acetylcholine receptor binding |
| publisher |
Nature Portfolio |
| publishDate |
2017 |
| url |
https://doaj.org/article/c37c2be9b26744c9a2ac701ad38d5c98 |
| work_keys_str_mv |
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