serum protein delipidation question

D.M. Bunk dmb618 at enh.nist.gov
Tue Mar 14 16:01:06 EST 1995


Does anyone have any "tried-and-true" methods (or 
references to methods) for defatting serum proteins?

I've been trying to isolate albumin from serum on a 
microscale using antibody affinity chromatography.  
While I have been able to extract the albumin, the 
albumin retains its complement of bound small 
lipophilic molecules (which interfere with the 
quantitation I ultimately want to do).

I have tried to defat the extracted albumin using the 
standard free fatty acid extraction solvent of 
chloroform/MeOH (2:1).  The problem here is that the 
MeOH precipitates the protein which settles on the 
chloroform-water interface, making it difficult to 
recover.

Does anyone have any suggestions?

-David




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