Rapid and Efficient Enrichment of Snake Venoms from Human Plasma Using a Strong Cation Exchange Tip Column to Improve Snakebite Diagnosis
Snake envenomation is a serious public health issue in many tropical and subtropical countries. Accurate diagnosis and immediate antivenom treatment are critical for effective management. However, the venom concentration in the victims’ plasma is usually low, representing one of the bottlenecks in d...
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doaj-5a6a74c944a1444fa9d0a179477cd5192021-02-14T00:00:17ZengMDPI AGToxins2072-66512021-02-011314014010.3390/toxins13020140Rapid and Efficient Enrichment of Snake Venoms from Human Plasma Using a Strong Cation Exchange Tip Column to Improve Snakebite DiagnosisChien-Chun Liu0Ya-Han Yang1Yung-Chin Hsiao2Po-Jung Wang3Jo-Chuan Liu4Chien-Hsin Liu5Wen-Chin Hsieh6Chih-Chuan Lin7Jau-Song Yu8Molecular Medicine Research Center, Chang Gung University, Taoyuan 333, TaiwanSchool of Medicine, College of Medicine, Chang Gung University, Taoyuan 333, TaiwanMolecular Medicine Research Center, Chang Gung University, Taoyuan 333, TaiwanMolecular Medicine Research Center, Chang Gung University, Taoyuan 333, TaiwanGraduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 333, TaiwanCenter for Research, Diagnostics and Vaccine Development of Centers for Disease Control, Ministry of Health and Welfare, Taipei 10050, TaiwanCenter for Research, Diagnostics and Vaccine Development of Centers for Disease Control, Ministry of Health and Welfare, Taipei 10050, TaiwanDepartment of Emergency Medicine, Chang Gung Memorial Hospital at Linkou, Taoyuan 333, TaiwanMolecular Medicine Research Center, Chang Gung University, Taoyuan 333, TaiwanSnake envenomation is a serious public health issue in many tropical and subtropical countries. Accurate diagnosis and immediate antivenom treatment are critical for effective management. However, the venom concentration in the victims’ plasma is usually low, representing one of the bottlenecks in developing clinically applicable assays for venom detection and snakebite diagnosis. In this study, we attempted to develop a simple method for rapid enrichment of venom proteins from human plasma to facilitate detection. Our experiments showed that several major protein components of both <i>Naja atra</i> (<i>N. atra</i>) and <i>Bungarus multicinctus</i> (<i>B. multicinctus</i>) venoms have higher isoelectric point (pI) values relative to high-abundance human plasma proteins and could be separated via strong cation exchange–high-performance liquid chromatography (SCX-HPLC). Based on this principle, we developed an SCX tip column-based protocol for rapid enrichment of <i>N. atra</i> and <i>B. multicinctus</i> venom proteins from human plasma. Application of liquid chromatography-tandem mass spectrometry (LC-MS/MS) led to the identification of cytotoxin and beta-bungarotoxin as the major proteins enriched by the SCX tip column in each venom sample. The entire process of venom enrichment could be completed within 10–15 min. Combination of this method with our previously developed lateral flow strip assays (rapid test) significantly enhanced the sensitivity of the rapid test, mainly via depletion of the plasma protein background, as well as increase in venom protein concentration. Notably, the SCX tip column-based enrichment method has the potential to efficiently enrich other <i>Elapidae</i> snake venoms containing proteins with higher pI values, thereby facilitating venom detection with other assays. This simple and rapid sample preparation method should aid in improving the clinical utility of diagnostic assays for snakebite.https://www.mdpi.com/2072-6651/13/2/140snakebitevenomstrong cation exchange chromatographylateral flow stripclinical diagnostic assay |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Chien-Chun Liu Ya-Han Yang Yung-Chin Hsiao Po-Jung Wang Jo-Chuan Liu Chien-Hsin Liu Wen-Chin Hsieh Chih-Chuan Lin Jau-Song Yu |
spellingShingle |
Chien-Chun Liu Ya-Han Yang Yung-Chin Hsiao Po-Jung Wang Jo-Chuan Liu Chien-Hsin Liu Wen-Chin Hsieh Chih-Chuan Lin Jau-Song Yu Rapid and Efficient Enrichment of Snake Venoms from Human Plasma Using a Strong Cation Exchange Tip Column to Improve Snakebite Diagnosis Toxins snakebite venom strong cation exchange chromatography lateral flow strip clinical diagnostic assay |
author_facet |
Chien-Chun Liu Ya-Han Yang Yung-Chin Hsiao Po-Jung Wang Jo-Chuan Liu Chien-Hsin Liu Wen-Chin Hsieh Chih-Chuan Lin Jau-Song Yu |
author_sort |
Chien-Chun Liu |
title |
Rapid and Efficient Enrichment of Snake Venoms from Human Plasma Using a Strong Cation Exchange Tip Column to Improve Snakebite Diagnosis |
title_short |
Rapid and Efficient Enrichment of Snake Venoms from Human Plasma Using a Strong Cation Exchange Tip Column to Improve Snakebite Diagnosis |
title_full |
Rapid and Efficient Enrichment of Snake Venoms from Human Plasma Using a Strong Cation Exchange Tip Column to Improve Snakebite Diagnosis |
title_fullStr |
Rapid and Efficient Enrichment of Snake Venoms from Human Plasma Using a Strong Cation Exchange Tip Column to Improve Snakebite Diagnosis |
title_full_unstemmed |
Rapid and Efficient Enrichment of Snake Venoms from Human Plasma Using a Strong Cation Exchange Tip Column to Improve Snakebite Diagnosis |
title_sort |
rapid and efficient enrichment of snake venoms from human plasma using a strong cation exchange tip column to improve snakebite diagnosis |
publisher |
MDPI AG |
series |
Toxins |
issn |
2072-6651 |
publishDate |
2021-02-01 |
description |
Snake envenomation is a serious public health issue in many tropical and subtropical countries. Accurate diagnosis and immediate antivenom treatment are critical for effective management. However, the venom concentration in the victims’ plasma is usually low, representing one of the bottlenecks in developing clinically applicable assays for venom detection and snakebite diagnosis. In this study, we attempted to develop a simple method for rapid enrichment of venom proteins from human plasma to facilitate detection. Our experiments showed that several major protein components of both <i>Naja atra</i> (<i>N. atra</i>) and <i>Bungarus multicinctus</i> (<i>B. multicinctus</i>) venoms have higher isoelectric point (pI) values relative to high-abundance human plasma proteins and could be separated via strong cation exchange–high-performance liquid chromatography (SCX-HPLC). Based on this principle, we developed an SCX tip column-based protocol for rapid enrichment of <i>N. atra</i> and <i>B. multicinctus</i> venom proteins from human plasma. Application of liquid chromatography-tandem mass spectrometry (LC-MS/MS) led to the identification of cytotoxin and beta-bungarotoxin as the major proteins enriched by the SCX tip column in each venom sample. The entire process of venom enrichment could be completed within 10–15 min. Combination of this method with our previously developed lateral flow strip assays (rapid test) significantly enhanced the sensitivity of the rapid test, mainly via depletion of the plasma protein background, as well as increase in venom protein concentration. Notably, the SCX tip column-based enrichment method has the potential to efficiently enrich other <i>Elapidae</i> snake venoms containing proteins with higher pI values, thereby facilitating venom detection with other assays. This simple and rapid sample preparation method should aid in improving the clinical utility of diagnostic assays for snakebite. |
topic |
snakebite venom strong cation exchange chromatography lateral flow strip clinical diagnostic assay |
url |
https://www.mdpi.com/2072-6651/13/2/140 |
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