Purification of purple coneflower root hemagglutinin by affinity chromatography

Abstract

Lecitins or hemagglutinins are non-immune origin glycoproteins, which can bind carbohydrate structures in specific and reversible manner. This particular feature enables lectin purification by affinity chromatography from complex protein samples. Proteins from Echinerea purpurea L. (Moench) roots haven't been investigated precisely, even though this herb is widely used in traditional medicine. Aim of experiment: To develop hemagglutinin from purple coneflower root purification by affinity chromatography. Experiment tasks: 1. Define carbohydrate affinity to hemagglutinins in purple coneflower root protein extracts; 2. Modify affinity matrix for lectin purification from purple coneflower root protein mixture; 3. Purity hemagglutinins from purple coneflower roots. Materials and methods: 1. To ascertain carbohydratte specificity of hemagglutinin component in crude purple coneflower root extract was determined by hemagglutination inhibition reaction in V bottomed microtiter plates. Twofold serial dilutions of crude protein extract were made and the sample was mixed and incubated for 1 h with equal volume of carbohydrate samples: 3 pentoses: D-arabinose, D-ribose, D-xylose, 5 hexoses and their derivates: D-galactose, D-mannose, D-glucose, Acetyl glucosamine, N-Acetyl glucosamine, 2 disaccharides D-lactose, D-maltose. [...]Botanikos sodasLietuvos sveikatos mokslų universitetas. Medicinos akademijaVilniaus universitetas. Biotechnologijos institutasVytauto Didžiojo universiteta