【正文】
rd Ion Exchange charge a) Mixture of proteins is applied to the negatively charged column resin – balanced by Na+ ions (red) b)ve and neutral proteins pass in the void volume, +ve proteins stick c) Excess of Na+ ions is added d)+ve proteins are eluted Could have used change in pH to elute the proteins CATION EXCHANGE Affinity Chromatography ? Uses specific binding property to attach protein to column ? Increase concentration of ligand to elute the protein ? Examples: – antibody columns monoclonal antibody attached to column – glutathione column – binds glutathioneStransferase on target 工業(yè)規(guī)模的色譜分離流程 色譜分離設(shè)備 玻璃色譜分離柱 色譜分離設(shè)備 帶熱夾套分離柱 反轉(zhuǎn)式分離柱 196。KTAexplorer 196。KTApurifier 196。KTAFPLC 色譜分離設(shè)備 色譜分離設(shè)備 實驗室層析和大規(guī)模層析的分別 實驗室 大規(guī)模生產(chǎn) 分辨率 越多峰越好 要求純度 種類 提高分辨率 純度 定性 增加 增加 樣品量 產(chǎn)量 (生產(chǎn)力 ) (峰越少越好 ) 上樣速度 不注重 最大樣品處理速度 次 /h 生產(chǎn)力 =g/ml 膠 /h 線性放大 固定以下條件: 1. 相同線行流速 2. 相同緩沖液 3. 相同填料 4