Phage-display of immunoglobulin heavy chain libraries
AssigneeThe United States of America as represented by the Department of Health and Human Services
InventorMary Jo Mulligan-Kehoe
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Abstract
One aspect of the invention relates to a phage-display library that expresses single-chain recombinant binding proteins. Inserts in the library comprise immunoglobulin heavy chain framework regions flanking highly divergent, synthetically produced hypervariable regions. A second aspect of the invention relates to the use of single-chain recombinant binding proteins to inhibit the activity of an intracellular constituent. In the exemplary case presented, the activity of intracellular glucose-6-phosphate dehydrogenase was inhibited by intracellular expression of a cloned single-chain recombinant binding protein.
Claims4
A phage-display library, comprising:
- a plurality of recombinant phage, each of said recombinant phage including an M13-derived expression vector having incorporated therein a polynucleotide coding for a polypeptide a protein encompassing a region spanning from upstream of an immunoglobulin heavy chain CDRI to a position downstream of CDRIII, wherein a DNA sequence encoding either CDRI or CDRIII is replaced by a randomly ordered DNA sequence, and wherein a fusion protein comprising the polypeptide encoded by said polynucleotide is expressed in the absence of an immunoglobulin light chain protein or portions thereof on an outer surface of recombinant phage of the library.
A phage display library comprising a plurality of recombinant phage, each of said recombinant phage including an M13-derived expression vector having incorporated therein a polynucleotide coding for a polypeptide encompassing a region spanning from upstream of an immunoglobulin heavy chain CDRI to a position downstream of CDRIII, wherein a DNA sequences encoding CDRI and CDRIII are each replaced by randomly ordered DNA sequences, and wherein a fusion protein comprising the polypeptide encoded by said polynucleotide is expressed in the absence of an immunoglobulin light chain polypeptide or portions thereof on an outer surface of recombinant phage of the library.
An isolated phage clone comprising:
- an M13-derived expression vector having incorporated therein a polynucleotide coding for a polypeptide encompassing a region spanning from upstream of an immunoglobulin heavy chain CDRI to a position downstream of CDRIlI, wherein a DNA sequence encoding CDRI or a DNA sequence encoding CDRIII is replaced by a randomly ordered DNA sequence, and wherein a fusion protein comprising the polypeptide encoded by said polynucleotide is expressed in the absence of an immunoglobulin light chain polypeptide or portions thereof on an outer surface of the phage clone.
Dependencies are inferred from the claim text.