Interference No. 102,572
Perry, ¶ 7 (CR-23-24), and protease could be removed by DEAE ion exchange
chromatography. Perry, ¶ 8 (CR-24).
(21) The strategy to reconstitute immunoglobulin chains also included the
comparison of the refolding results of heavy and light chains from the anti-CEA antibodies
produced from hybridoma cells. Perry, ¶ 10. The antibodies were said to be supplied by
18
Cabilly. Optimal conditions were determined and used. Perry, ¶ 10.
(22) Perry stated that later they found that they could isolate the heavy and light
chains from both single and cotransformed bacteria by guanidine solubilization of refractile
body preparations without any further purification by column chromatography. These
samples could then be used directly in the refolding reactions. The final conditions
involved a mixture of sulfitolized heavy chain and an extract of light chain cells. The chains
were reconstituted using the same conditions as the optimal conditions for the
reconstitution of antibody chains produced in hybridoma cells. Perry, ¶ 11 (CR-26) .
(23) Wetzel testified that he recorded in his notebook the results of a Western blot
of SDS-PAGE run by Perry. He testified that from the blot they noted production of heavy
and light chain protein product in the co-transformed E.coli cells and that they were able to
estimate the level of expression (%) from cell paste from Mumford. Wetzel testified that the
results were used to calculate the theoretical maximum possible yield which were in turn
18The Cabilly et al. brief (page 19, last 5 lines-page 20, line 1) lists a number of
conditions not identified by Perry in her testimony. id.
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