层析计数相关培训课件.pdf

层析基本原理及工艺的建立 Chromatoggrapphyy Techniqques IIn ordder to purifify proteiins, peptidides andd nuclleiic acidids we need powerful techniques. Content Principles of Operation TermsTerms ParametersParameters Advantages Disadvantages of different techniquestechniques Optimization of separation techniques PrinciplesPrinciples ofof OperationOperation forfor Chromatography Techniques Gel Ion Hydrophobic Affinity Reversed Filtration Exchange Interaction Phase Impportant terms pparameters ResolutionResolution Efficiency Selectivity Symmetry Resolution: the power to separate peaks Maximum number of peaks in gel filtration ca 15 22 MMaxiimum numbber off peakks iin RPCRPC 150150 4 0 2 8 3 1 6 6 0 10 7 20 ml 5 4 2 V0 Vt 0 0 5 10 15 20 25 0 200 400 ml Remember: we are not pp urifyfy ingg pp eaks, but pp roteins pp epp tides ! Resolution factor R s Rs = 0.6 Resolution = Peak separation PeakPeak widthwidth atat ½½ heightheight Rs = 1