C - Chemistry – Metallurgy – 12 – P
Patent
C - Chemistry, Metallurgy
12
P
C12P 21/08 (2006.01) C12N 5/18 (2006.01) G01N 33/573 (2006.01) G01N 33/574 (2006.01) G01N 33/577 (2006.01)
Patent
CA 2108533
A monoclonal antibody against cytosolic nucleoside diphosphate kinase (NDP-kinase: EC 2.7.4.6) from HL60 cells has been produced. The monoclonal antibody solely recognizes the active oligomeric form of NDP-kinase and does not inhibit NDP-kinase activity. Reduction and alkylation of NDP-kinase oligomer performed under denaturing conditions fully destroyed the antigenic site recognized by the monoclonal antibody. Western blot analysis after non-denaturing polyacrylamide gel electrophoresis (PAGE) revealed that a single protein oligomer is recognized by the MAb. The protein band showed the same electrophoretic mobility of pure cytosolic NDP-kinase detected by a specific NDP-kinase enzymic staining assay performed after nondenaturing PAGE. Collectively, the results suggest that the MAb recognizes the native form of NDP-kinase. Searching for the reactivity of the MAb against membrane-bound NDP-kinase it was found that the MAb selectively reacts with a cell-membrane component of monocytes (93%), granulocytes (97%) and 14% of lymphocytes. Two color immunofluorescence showed that the MAb reacted only with CD19+ B cells (93% of CD19+ B cells), which suggest that NDP-kinase may play an important role in normal B lymphocyte function.
Anderson J. Wayne
Her Majesty The Queen In Right Of Canada As Represented By The Minister Of Health
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