MAA/SNA Lectins

Ian McFarlane imcfarlane at ninewells.dundee.ac.uk
Wed Sep 27 11:29:24 EST 1995


I am attempting to purify a secreted/cell surface protein from a transformed 
cell line using lectin-affinity chromatography with lectins specific for 
sialic acid. However, no protein appears to be sticking to the columns. The 
columns are 0.5.x5cm packed with 1ml MAA/SNA gel (3mg protein). I have tried 
loading the samples in DMEM and in PBS, and have eluted either by gravity or 
at 0.2ml/min with PBS followed by either 0.1M glycine pH3.5 or 20mM 
ethylenediamine. When this failed I attempted to use standard glycoproteins to 
check if the columns were OK both fetuin for the MAA and transferrin for the 
SNA failed to adhere in significant amounts. My secreted/cell surface protein 
is reactive with the lectins after Western blotting  using DIG-labelled 
lectins in the Boehringer Glycan Differentiation kit. Does anyone have any 
ideas as to what the problem may be? 

                                                  Thanks Ian Mc



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