Appeal No. 96-2137
Application 07/668,920
radiolabeled anti-mouse IgG may be used to quantitate the amount of
bound mouse antibody. Direct or indirect immunofluorescence screening
techniques may also be used. Specificity for insoluble intracellular
antigens may be determined by comparing the amount of antibody bound
to cell ghosts with that bound to intact cells.
The specification also describes the manner in which hybridomas which produce
monoclonal antibodies to nuclear antigens were prepared. As set forth in Example 1
(the paragraph bridging pages 16-17 of the specification):
In order to generate hybridomas producing monoclonal antibody to
nuclear antigens, eight human malignant lymphoma and leukemia cell
lines were used as a source of antigens. These include the EBV-positive
nonproducer Raji and producer AG876 African Burkitt’s lymphoma cell
lines; the T-cell acute lymphoblastic leukemia CEM cell line; the IgE
secreting multiple myeloma U-266 cell line; the erythroleukemia K562 cell
line; and the histiocytic type SU-DHL-1 and U-937 and B-cell type SU-
DHL-4 diffuse histiocytic lymphoma cell lines. In addition to these
cultures, normal peripheral blood lymphocytes pooled from several
individuals and separated by the ficoll-hypaque technique were used
alone and after four days of stimulation with 5ug/ml of Pokeweed mitogen.
Example 2 of the specification discusses the results obtained when certain monoclonal
antibodies were screened. As set forth at page 20 of the specification, the monoclonal
antibodies screened in this example were selected “from a library of monoclonal
antibodies to intracellular antigens that includes the antibodies produced by the
hybridomas of Example 1.” The selected antibodies were screened using large cell
lymphoma cells (SU-DHL-2) and adenocarcinoma lung cancer cells (A549). The
results of the screening are set forth at Table 1 of the specification as follows:
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Last modified: November 3, 2007