PREPARATION AND PRELIMINARY APPLICATION OF MONOCLONAL ANTIBODIES AGAINST RECOMBINANT HUMAN GRANULOCYTE COLONY STIMULATING FACTOR
Chen X, Fang XD*, Wang XG*, Wang XN*.
Institute of Biotechnology, Sun Yat Sen University of Medical Sciences, Guangzhou and *Institute of Molecular Immunology, First Military Medical University, Guangzhou, China
The study overcame the problems of weakly immunogenic granulocyte colony-stimulating factor (G-CSF) to mouse by utilizing the typhoid bacteria body to produce 3 murine hybridoma clones of monoclonal antibodies against recombinant human G-CSF. The 3 hybridoma clones were named 1H11, 2B3 and 2C3. These antibodies were all IgG2a. Western blot was applied to test the specificity of antibodies against G-CSF. The McAb additivity test and McAb competitive test showed the 2B3 and 2C3 recognized the same epitope of G-CSF while 1H11 recognized the other epitope of G-CSF. The neutralization trial of G-CSF indicated 2B3 and 2C3 were capable of negating the proliferative activity of G-CSF in NFS-60 cells. The double antibody sandwich ELISA was established by using these McAbs to detect G-CSF levels in human serum. The sensitivity to G-CSF was 340 pg/ml. G-CSF levels of human serum samples were also evaluated. It was found that no positivity was encountered in normal serum samples. However, G-CSF was detected in 91.6% of patients with bacterial infection and in 25-35.7% of leukemia patients. The study provided a simple, rapid and specific assay of G-CSF in human serum. The assay overcame the flaw of myeloid progenitor cell assay and NFS-60 cell assay, which were tedious and time-consuming.
Copyright 1997 Hong Kong Medical Technology Association .
All rights reserved.