Multipurpose HTS Coagulation Analysis: Assay Development and Assessment of Coagulopathic Snake Venoms
Coagulation assays currently employed are often low throughput, require specialized equipment and/or require large blood/plasma samples. This study describes the development, optimization and early application of a generic low-volume and high-throughput screening (HTS) assay for coagulation activity...
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| Format: | Article |
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MDPI AG
2017-11-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/9/12/382 |
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| author | Kristina B. M. Still Randjana S. S. Nandlal Julien Slagboom Govert W. Somsen Nicholas R. Casewell Jeroen Kool |
| author_facet | Kristina B. M. Still Randjana S. S. Nandlal Julien Slagboom Govert W. Somsen Nicholas R. Casewell Jeroen Kool |
| author_sort | Kristina B. M. Still |
| collection | DOAJ |
| description | Coagulation assays currently employed are often low throughput, require specialized equipment and/or require large blood/plasma samples. This study describes the development, optimization and early application of a generic low-volume and high-throughput screening (HTS) assay for coagulation activity. The assay is a time-course spectrophotometric measurement which kinetically measures the clotting profile of bovine or human plasma incubated with Ca2+ and a test compound. The HTS assay can be a valuable new tool for coagulation diagnostics in hospitals, for research in coagulation disorders, for drug discovery and for venom research. A major effect following envenomation by many venomous snakes is perturbation of blood coagulation caused by haemotoxic compounds present in the venom. These compounds, such as anticoagulants, are potential leads in drug discovery for cardiovascular diseases. The assay was implemented in an integrated analytical approach consisting of reversed-phase liquid chromatography (LC) for separation of crude venom components in combination with parallel post-column coagulation screening and mass spectrometry (MS). The approach was applied for the rapid assessment and identification of profiles of haemotoxic compounds in snake venoms. Procoagulant and anticoagulant activities were correlated with accurate masses from the parallel MS measurements, facilitating the detection of peptides showing strong anticoagulant activity. |
| first_indexed | 2024-04-14T00:59:07Z |
| format | Article |
| id | doaj.art-3e6147aa818048239f9463057448c71d |
| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-04-14T00:59:07Z |
| publishDate | 2017-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-3e6147aa818048239f9463057448c71d2022-12-22T02:21:27ZengMDPI AGToxins2072-66512017-11-0191238210.3390/toxins9120382toxins9120382Multipurpose HTS Coagulation Analysis: Assay Development and Assessment of Coagulopathic Snake VenomsKristina B. M. Still0Randjana S. S. Nandlal1Julien Slagboom2Govert W. Somsen3Nicholas R. Casewell4Jeroen Kool5Division of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsDivision of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsDivision of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsDivision of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsAlistair Reid Venom Research Unit, Parasitology Department, Liverpool School of Tropical Medicine, Liverpool L3 5QA, UKDivision of BioAnalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, De Boelelaan 1085, 1081 HV Amsterdam, The NetherlandsCoagulation assays currently employed are often low throughput, require specialized equipment and/or require large blood/plasma samples. This study describes the development, optimization and early application of a generic low-volume and high-throughput screening (HTS) assay for coagulation activity. The assay is a time-course spectrophotometric measurement which kinetically measures the clotting profile of bovine or human plasma incubated with Ca2+ and a test compound. The HTS assay can be a valuable new tool for coagulation diagnostics in hospitals, for research in coagulation disorders, for drug discovery and for venom research. A major effect following envenomation by many venomous snakes is perturbation of blood coagulation caused by haemotoxic compounds present in the venom. These compounds, such as anticoagulants, are potential leads in drug discovery for cardiovascular diseases. The assay was implemented in an integrated analytical approach consisting of reversed-phase liquid chromatography (LC) for separation of crude venom components in combination with parallel post-column coagulation screening and mass spectrometry (MS). The approach was applied for the rapid assessment and identification of profiles of haemotoxic compounds in snake venoms. Procoagulant and anticoagulant activities were correlated with accurate masses from the parallel MS measurements, facilitating the detection of peptides showing strong anticoagulant activity.https://www.mdpi.com/2072-6651/9/12/382snake venomsblood coagulationcoagulation bioassayhigh-throughput screeningmass spectrometryliquid chromatography |
| spellingShingle | Kristina B. M. Still Randjana S. S. Nandlal Julien Slagboom Govert W. Somsen Nicholas R. Casewell Jeroen Kool Multipurpose HTS Coagulation Analysis: Assay Development and Assessment of Coagulopathic Snake Venoms Toxins snake venoms blood coagulation coagulation bioassay high-throughput screening mass spectrometry liquid chromatography |
| title | Multipurpose HTS Coagulation Analysis: Assay Development and Assessment of Coagulopathic Snake Venoms |
| title_full | Multipurpose HTS Coagulation Analysis: Assay Development and Assessment of Coagulopathic Snake Venoms |
| title_fullStr | Multipurpose HTS Coagulation Analysis: Assay Development and Assessment of Coagulopathic Snake Venoms |
| title_full_unstemmed | Multipurpose HTS Coagulation Analysis: Assay Development and Assessment of Coagulopathic Snake Venoms |
| title_short | Multipurpose HTS Coagulation Analysis: Assay Development and Assessment of Coagulopathic Snake Venoms |
| title_sort | multipurpose hts coagulation analysis assay development and assessment of coagulopathic snake venoms |
| topic | snake venoms blood coagulation coagulation bioassay high-throughput screening mass spectrometry liquid chromatography |
| url | https://www.mdpi.com/2072-6651/9/12/382 |
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