Anti-ngf antibodies for the treatment of various disorders
a technology of anti-ngf antibodies and disorders, applied in the field of anti-ngf antibodies, can solve the problems of significant public health problems such as chronic inflammatory conditions such as these, and the current therapies for treating these conditions are severely limited
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example 1
Production And Characterization of Anti-NGF Monoclonal Antibodies
[0137]A. Production of Anti-NGF Monoclonal Antibodies
[0138]This example illustrates preparation of monoclonal antibodies that can specifically bind human NGF (hNGF). Techniques for producing monoclonal antibodies are well known in the art and are described, for instance, in Kohler and Milstein, Nature 256:495-497 (1975). The experiments described in Examples 1 and 2 are further described in Hongo et al., Hybridoma 19:215-227.
[0139]A panel of 23 murine monoclonal antibodies to hNGF was developed by a method analogous to that described in Hongo et al., Hybridoma 14:253-260. Briefly, Balb / c mice (Charles River Laboratories, Wilmington, Del.) were immunized with human NGF in Ribi adjuvant (Ribi Immunochem Research, Inc., Hamilton, Mo.). Splenocytes from the mouse demonstrating the highest titer of antibody to immobilized NGF were fused with mouse myeloma cells (X63.Ag8.653; American Type Culture Collection, Rockville, Md.)...
example 2
Determination of Neutralizing Activity of Anti-NGF Monoclonal Antibodies
[0152]The observation that some epitopes mapped in regions previously shown to be significant in the interaction of NGF with TrkA and / or p75 suggested that the corresponding MAbs might also block one or both of these interactions.
[0153]A. 125I-hNGF Binding Assay
[0154]To evaluate the possibility that one or more of the anti-NGF MAbs might block NGF binding to TrkA and / or p75, the binding of 125I-hNGF to the TrkA-IgG receptor immunoadhesin was measured in the presence of anti-NGF monoclonal antibodies.
[0155]Briefly, 125I-hNGF was prepared using a modification of the soluble lactoperoxidase method originally described by Marchalonis (Marchalonis, Biochem J., 113:299-305 (1969)). The final reaction mixture was fractionated over a pD-10 Sephadex 6-25 size exclusion column (Pharmacia, Uppsala, Sweden) and stored at 4° C. Microtiter plates (Nunc, Maxisorb) were coated overnight at 4° C. with purified rabbit anti-human ...
example 3
The Effect of Anti-NGF On the Immune Response
[0169]Mice were immunized subcutaneously with 10 μg ovalbumin on day 0 and allowed to recover. On day 40 following immunization animals were injected IP with 10 mg / kg anti-NGF antibody 911 or a control, isotype matched antibody. Two days later animals were boosted with ovalbumin. The immune response was measured by ELISA on serum samples taken on day 47. As can be seen in FIG. 10, there was no significant (p>0.05) difference in the immune response between the animals receiving anti-NGF antibody 911 and animals receiving the control antibody. However, there was a non-significant trend towards an increase in the immune response in animals treated with the anti-NGF monoclonal antibody.
[0170]By contrast, FIG. 11 shows that following immunization with chicken gamma-globulin, treatment with anti-NGF produces a non-significant trend toward a decrease in the immune response. Animals were immunized with 10 μg of chicken gamma-globulin on day 0 and...
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