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Affinity Chromatography is
the most powerful technique
but cannot be applied to all
systems
Mw (D)
● Vt = Vx + Vo , where Vt is the total column volume, Vx is volume
occupied by the stationary phase and Vo is void (remaining) volume
● The ratio of the elution and void volumes (Ve/Vo) varies approximately
linearly with log Mw
– Outliers on above plot tend to have highly asymmetric shapes (non-spherical)
Separated
proteins
Resolving
Gel pH 8.8
Positive
+ +
Glycine: H3N+CH2COO- H2NCH2COO- + H+