Appeal No. 2002-1547 Page 6
Application No. 09/259,434
chromatography on a “Sephacryl S-200 column (2.6 x 150 cm),” followed by
chromatography on a “Thiopropyl-Sepharose column.” See pages 10-11. When
the monomers were dimerized, the resulting compositions comprised no more
than 36.1% dimers. See pages 12-13.
Sirtori discloses purification of recombinant Apo AI-M by chromatography
on either a “Sephacryl S-200 column and/or an anti-apo AI-Sepharose column.”
Page 12. The examiner has pointed to no other disclosure in Sirtori that would
teach or suggest other purification methods to those skilled in the art. Nor has
the examiner presented evidence that either of the disclosed methods would
purify Apo AI-M dimers to at least 90% homogeneity. The evidence, in fact, is to
the contrary. The instant specification discloses that Sephacryl S-200
chromatography does not result in a purified dimer preparation, but rather
produces monomers that must be oxidized to produce even a reasonable (36%)
amount of dimers.
In addition, Sirtori analyzed the purified Apo AI-M by circular dichroism
(CD) spectroscopy and found that it had an α-helix content of 43%, lower than
the 52% found for normal Apo AI. See page 12. The instant specification, on the
other hand, discloses that purified Apo AI-M dimers have an α-helix content of
52.2% to 57.8%, depending on protein concentration. See page 22. The CD
spectroscopy data provide further evidence that Apo AI-M composition disclosed
by Sirtori does not contain Apo AI-M dimers purified to at least 90%
homogeneity.
Page: Previous 1 2 3 4 5 6 7 8 9 Next
Last modified: November 3, 2007