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US 5,633,130 AGranted
Buffer system for increasing seroconversion efficiency
AssigneeThe Curators of the University of Missouri
InventorsDaniel S. Smith, John C. Walker
Life of this patent
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Abstract
A method of increasing efficiency of deantigenation of blood group epitopes on erythrocytes (seroconversion) by exoglycosidases utilizing a step of performing the deantigenation in a zwitterionic buffer. The method provides a buffer system and exoglycosidase that utilizes a twenty to one hundred fold lower total enzyme mass than the prior art methods.
Claims7
A method of preparing deantigenated erythrocytes including the steps of
- isolating erythrocytes of a defined blood type;
- suspending the isolated erythrocytes in zwitterionic buffer wherein the zwitterionic buffer consists essentially of 0.1 to 20 mM Na titrate, 220 to 440 mM glycine or alanine and 0.01 to 30 mg ml.sup.-1 albumin at pH 5.8;
- adding a multimeric eucaryotic exoglycosidase with multiple subunits selected to act on the isolated erythrocytes;
- incubating at 24.degree. C. for between one and two hours;
- and
- washing in phosphate buffered saline.
3 more elements
A deantigenation buffer consisting of 0.1 to 20 mM Na citrate, 220 to 440 mM glycine and 0.1 to 30 mg ml.sup.-1 albumin at pH 5.8.
Dependencies are inferred from the claim text.