期刊
BIOLOGICALS
卷 32, 期 1, 页码 37-47出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.biologicals.2003.12.002
关键词
G-CSF; glycosylation; serum; alpha 2-macroglobulin; biological activity
It has previously been reported that the biological activity of the human hematopoetic cytokine granulocyte colony stimulating factor (G-CSF) was reduced following incubation with human serum. The mechanism of action of serum has remained elusive although a number of possible mechanisms have been suggested including inactivation due to binding to the serum protein a(2)-macroglobulin (a(2)M) and degradation by serum proteases. The aim of this study was to clarify the conditions required by serum to reduce the biological activity of the cytokine and to define the mechanism involved. It has also been noted that G-CSF obtained from a CHO expression system (and therefore considered a glycosylated molecule) was resistant to serum inactivation unlike G-CSF obtained from an E coli expression system (considered to be non-glycosylated). We used an enzymatic approach to remove the carbohydrate residues from glycosylated G-CSF and tested this material for its stability in serum. We additionally used a mutated G-CSF lacking glycosylation sites. We concluded that glycosylation was important in protecting against serum inactivation. We observed that serum reduced the biological activity of non-glycosylated G-CSF in a dose, and temperature dependent manner and deduced that the mechanism of action was dependent upon a(2)M bound serum protease enzymes. (C) 2003 The International Association for Biologicals. Published by Elsevier Ltd. All rights reserved.
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