Fusion proteins to facilitate selection of cells infected with specific immunoglobulin gene recombinant vaccinia virus
View Patent ↗The present invention relates to a high efficiency method of expressing immunoglobulin molecules in eukaryotic cells. The invention is further drawn to a method of producing immunoglobulin heavy and light chain libraries, particularly using the trimolecular recombination method, for expression in eukaryotic cells. The invention further provides methods of selecting and screening for antigen-specific immunoglobulin molecules, and antigen-specific fragments thereof. The invention also provides kits for producing, screening and selecting antigen-specific immunoglobulin molecules. Finally, the invention provides immunoglobulin molecules, and antigen-specific fragments thereof, produced by the methods provided herein.
1. A fusion protein comprising from N-terminus to C-terminus:
(a) an immunoglobulin heavy chain variable region;
(b) a polypeptide segment consisting of the CH1 domain of human IgG1, or a polypeptide consisting of the full-length constant region of human IgG1; and
(c) amino acids 215 to 421 of SEQ ID NO:11;
wherein amino acids 215 to 421 of SEQ ID NO:11 are identical to amino acids 108 to 314 of A56R from the Western Reserve Vaccinia virus strain.
2. The fusion protein of claim 1 , further comprising a signal peptide for facilitating expression of the fusion protein on the surface of EEV.
3. The fusion protein of claim 1 , wherein the polypeptide segment of (b) consists of the CH1 domain of human IgG1 fused to amino acids 215 to 421 of SEQ ID NO: 11.
4. The fusion protein of claim 1 , wherein the polypeptide segment of (b) consists of the full-length constant region of human IgG1 fused to amino acids 215 to 421 of SEQ ID NO: 11.