Fast protein liquid chromatography for the purification of animal venoms.
In this paper a methodology is presented for the purification of toxins acting on neuronal potassium channels. These neurotoxins are ideal tools for studying the physiological functioning of ion channels, for classifying them and for exploring them in several neuronal regions. Separation procedures leading to the identification of neurotoxins almost always include gel filtration chromatography as well as other chromatographic techniques such as ion exchange and reversed phase chromatography. The use of a Superdex 30 prep grade gel filtration column, which has the advantage of superior resolution in comparison with traditionally used gel filtration column, which has the advantage of superior resolution in comparison with traditionally used gel filtration columns such as a Sephadex G-50 column, is reported here. The biological assay incorporates the use of Xenopus laevis oocytes, which express potassium channels. Screening of the collected venom fractions was performed by means of the whole-cell voltage clamp technique. The combination of these techniques represents a fast and efficient identification procedure in the search for new and selective neurotoxins for cloned channels and receptors.
Debont
T
T
Laboratory of Toxicology, Faculty of Pharmaceutical Sciences, University of Leuven, Belgium.
Daenens
P
P
Tytgat
J
J
D000818:Animals; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004546:Elapid Venoms / Q000302:isolation & purification* / Q000494:pharmacology; D005260:Female; D066298:In Vitro Techniques; D009498:Neurotoxins / Q000302:isolation & purification* / Q000494:pharmacology; D009865:Oocytes; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D014982:Xenopus laevis
D004546:Elapid Venoms; D009498:Neurotoxins; D015221:Potassium Channels