Immunoglobulin Chimeric Monomer-Dimer Hybrids
AssigneeSyntonix Pharmaceuticals, Inc.
InventorsRobert T. PETERS, Adam R. Mezo, Daniel S. Rivera, Alan J. Bitonti, James Stattel, Susan C. Low
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Abstract
The invention relates to a chimeric monomer-dimer hybrid protein wherein said protein comprises a first and a second polypeptide chain, said first polypeptide chain comprising at least a portion of an immunoglobulin constant region and a biologically active molecule, and said second polypeptide chain comprising at least a portion of an immunoglobulin constant region without the biologically active molecule of the first chain. The invention also relates to methods of using and methods of making the chimeric monomer-dimer hybrid protein of the invention.
Claims42
A chimeric protein comprising a first and second polypeptide chain, wherein said first chain comprises a biologically active molecule, and at least a portion of an immunoglobulin constant region and wherein said second chain comprises at least a portion of an immunoglobulin constant region without a biologically active molecule or immunoglobulin variable region.
-31. (canceled)
A chimeric protein comprising a first and second polypeptide chain
- a) wherein said first chain comprises a biologically active molecule, at least a portion of an immunoglobulin constant region, and a first domain having at least one specific binding partner;
- and
- b) wherein said second chain comprises at least a portion of an immunoglobulin without a biologically active molecule or immunoglobulin variable region and further comprising a second domain said second domain being a specific binding partner of said first domain.
-64. (canceled)
A method of making a biologically active chimeric protein comprising:
- a) transfecting a cell with a first DNA construct comprising a DNA molecule encoding a polypeptide comprising a biologically active molecule operatively linked to a second DNA molecule encoding at least a portion of an immunoglobulin constant region;
- b) transfecting the cell with a second. DNA construct comprising a DNA molecule encoding a polypeptide comprising at least a portion of an immunoglobulin constant region without a biologically active molecule or variable region of an immunoglobulin;
- c) culturing the cell under conditions such that the polypeptide encoded by said first DNA construct and said second DNA construct is expressed;
- and
- d) isolating dimers of a) and b) from said transfected cell.
-71. (canceled)
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-84. (canceled)
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-98. (canceled)
-130. (canceled)
-153. (canceled)
A method of making a chimeric protein comprising an Fc fragment of an immunoglobulin linked to a biologically active molecule, said method comprising
- a) transfecting a cell with a DNA construct comprising a DNA sequence encoding an Fc fragment of an immunoglobulin and a second DNA sequence encoding intein;
- b) culturing said cell under conditions such that the Fc fragment and intein is expressed;
- c) isolating said Fc fragment and intein from said cell;
- d) chemically synthesizing a biologically active molecule having an N terminal Cys;
- e) reacting the isolated intein Fc of c) with MESNA to generate a C terminal thio-ester;
- f) reacting the biologically active molecule of d) with the Fc of e) to make a chimeric protein comprising an Fc linked to a biologically active molecule.
-159. (canceled)
-194. (canceled)
A method of making a biologically active chimeric protein comprising:
- a) transfecting a first cell with a first DNA construct comprising a DNA molecule encoding a polypeptide comprising a biologically active molecule operatively linked to a second DNA molecule encoding at least a portion of an immunoglobulin constant region;
- b) transfecting a second cell with a second DNA construct comprising a DNA molecule encoding a polypeptide comprising at least a portion of an immunoglobulin constant region without a biologically active molecule or variable region of an immunoglobulin;
- c) culturing the cell of a) and b) under conditions such that the polypeptide encoded by said first DNA construct and said second DNA construct is expressed;
- and
- d) isolating dimers of a) and b) from said transfected cells.
Dependencies are inferred from the claim text.