Appeal No. 2002-1547 Page 3
Application No. 09/259,434
appears that these subjects may be “protected” from
atherosclerosis.
. . .
Another very specific feature of the Apo AI-M, is its capacity to form
dimers with itself and complexes with Apo AII, in both cases
because of the presence of the Cys residue. From studies of blood
fractions containing a mixture of Apolipoproteins, there were
indications, showing that the presence of dimers and complexes in
the circulation may be responsible for the increased elimination
half-life of these in the carriers, recently described in clinical
studies.
Id., pages 4-5.
The specification discloses recombinant production of Apo AI-M dimers.
(pages 14-21) and purification of the Apo AI-M dimers from the plasma of
subjects carrying the Apo AI-M mutation. See pages 10-13. The dimers were
purified directly from plasma by two steps of chromatography on a “Sephacryl S-
300 HR column (2.6 x 300 cm).” Page 10. The resulting dimer preparations are
disclosed to be “>98% pure.” Id.
The specification also discloses that Apo AI-M dimers can be made by
isolating Apo AI-M monomers, then converting them to dimers. See pages 10-
13. Monomers were purified by a single step of chromatography on a “Sephacryl
S-200 column (2.6 x 150 cm),” followed by chromatography on a “Thiopropyl-
Sepharose column” to separate normal Apo AI from Apo AI-M. See pages 10-
11. The Apo AI-M monomers were then converted to dimers by oxidation,
resulting in up to 36.1% dimer formation. See pages 12-13.
The plasma-purified dimers were characterized by several methods.
Circular dichroism spectroscopy showed that the purified dimers had an α-helix
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