Thông tin tài liệu


Title: Development of high-throughput screening assays for profiling snake venom Phospholipase A2 activity after high-resolution chromatographic fractionation
Authors: Still, Kristina B.M
Slagboom, Julien
Kidwai, Sarah
Keywords: Bothrops asper; Echis carinatus; Echis coloratus; Echis ocellatus, Oxyuranus scutellatus; Daboia russelii russelii
Issue Date: 2020
Publisher: Biochemical Journal
Abstract: Many organisms, ranging from plants to mammals, contain phospholipase A2 enzymes (PLA2s), which catalyze the production of lysophospholipids and fatty acid proinflammatory mediators. PLA2s are also common constituents of animal venoms, including bees, scorpions and snakes, and they cause a wide variety of toxic effects including neuro-, myo-, cyto-, and cardio-toxicity, anticoagulation and edema. The aim of this study was to develop a generic method for profiling enzymatically active PLA2s in snake venoms after chromatographic separation. For this, low-volume high-throughput assays for assessment of enzymatic PLA2 activity were evaluated and optimized. Subsequently, the assays were incorporated into a nanofractionation platform that combines high-resolution fractionation of crude venoms by liquid chromatography (LC) with bioassaying in 384-well plate format, and parallel mass spectrometric (MS) detection for toxin identification. The miniaturized assays developed are based on absorbance or fluorescence detection (respectively, using cresol red or fluorescein as pH indicators) to monitor the pH drop associated with free fatty acid formation by enzymatically active PLA2s. The methodology was demonstrated for assessment of PLA2 activity profiles of venoms from the snake species Bothrops asper, Echis carinatus, Echis coloratus, Echis ocellatus, Oxyuranus scutellatus and Daboia russelii russelii.
Description: Tài liệu này được phát hành theo giấy phép CC-BY - ND 4.0
URI: http://dlib.hust.edu.vn/handle/HUST/24805
Link item primary: https://www.biorxiv.org/content/10.1101/2020.01.20.912758v1
Appears in Collections:OER - Kỹ thuật hóa học; Công nghệ sinh học - Thực phẩm; Công nghệ môi trường
ABSTRACTS VIEWS

12

VIEWS & DOWNLOAD

9

Files in This Item:
Thumbnail
  • OER000000928.pdf
      Restricted Access
  • Nội dung
    • Size : 4,25 MB

    • Format : Adobe PDF



  • This item is licensed under a Creative Commons License Creative Commons