Appeal 2007-1161
Application 09/954,166
cells and had a high nanomolar affinity for T-cell clones which could be
measured by a direct binding assay (Dal Porto, p. 6671, col. 1 (Abstract);
p. 6672, col. 1; Answer 5).
10. Dal Porto’s genetically engineered divalent class I MHC molecule
comprises: 1) the extracellular MHC binding (H-2Kb) domain (α1, α2, and
α3) joined to 2) the variable (V) region of IgG heavy chain, forming a
chimeric protein (Dal Porto, p. 6672, cols. 1-2; Fig. 1B).
11. To make this molecule, the gene encoding the chimeric protein (H-
2Kb/IgG) was expressed in a cell line (J558L) which also expresses an
immunoglobulin light chain (Dal Porto, p. 6672, col. 1) and beta2-
microglobulin.
12. The resulting molecule was purified from culture. Analysis showed that
the molecule was divalent (containing two MHC binding sites), consisting
“of dimers of a complex composed of chimeric heavy chain,
immunoglobulin light chain, and β2-microglobulin (Fig. 1B)” (Dal Porto, p.
6673, col. 2). The structure of Dal Porto’s molecule is reproduced in the
figure below:
Dal Porto’s figure shows a protein complex comprising chimeric
heavy chain, immunoglobulin light chain, and β2-microglobulin.
5
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