C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 5/08 (2006.01) A61K 35/28 (2006.01) C07K 16/28 (2006.01) C12N 5/06 (2006.01) C12Q 1/24 (2006.01) C12S 3/00 (2006.01) G01N 33/569 (2006.01) A61K 35/12 (2006.01)
Patent
CA 2108565
2108565 9218643 PCTABS00017 A method for recovering viable cells having surface protein determinants having portions rich in O-glycosylated carbohydrates is provided. The proteins have determinants which are substrate sensitive to a P. haemolytica derived neutralmetallo-glycoprotease. The glycoprotease has highly restricted specificity for cleaving solely from the cell surfaces the O-glycosylated protein portions having the determinants while retaining cell viability. The process comprises: i) contacting the cells in solution with affinity matrices which bind specifically to one or more of the determinants which are part of the glycoprotease sensitive substrates, the affinity matrices are allowed to bind to the determinants on the cells, ii) separating the cells to which the affinity matrices are bound from any remaining matter in the solution, the affinity matrices having sufficient binding affinity for the determinants to remain bound to the determinants during the separation, iii) contacting the glycoprotease with the separated cells in sufficient concentration and duration to cleave solely the protein substrate portions having the determinants and bound by the affinity matrices and thereby release the cells from the affinity matrices, the released cells retain viability due to the restricted specificity of the glycoprotease cleaving solely O-glycosylated protein portions having the determinants, and iv) recovering the released cells.
Mellors Alan
Sutherland Robert D.
Bereskin & Parr
The Toronto Hospital
University Of Guelph
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