Synthesis of protein with an identification peptide, and hybrid

Details Synthesis of protein with an identification peptide, and hybrid Synthesis of protein with an identification peptide, and hybrid

1987-07-23

1988-11-01

Foelak, Morton

Chemistry: natural resins or derivatives; peptides or proteins;

Peptides of 3 to 100 amino acid residues

8 to 10 amino acid residues in defined sequence

530330, 530350, 435 68, A61K 3702, C12P 2100

Patent

active

047821370

ABSTRACT:
A hybrid polypeptide composed of an identification peptide and a desired functional protein are produced by recombinant DNA techniques. A DNA expression vector is constructed that includes segments of DNA coding for the identification peptide and the desired functional protein. The identification peptide consists of a highly antigenic N-terminal portion and a C-terminal linking portion that connects the identification peptide to the N-terminal of the functional protein. The linking portion of the identification peptide is cleavable at a specific amino acid residue adjacent the functional protein by use of a sequence specific proteolytic enzyme or chemical proteolytic agent. The hybrid polypeptide expressed by the host cells transformed by the cloning vector is removed therefrom and purfied by affinity chromatography techniques by use of an immobilized ligand specific to the antigenic portion of the identification peptide. The protein is then cleaved from the isolated hybrid polypeptide with an appropriate proteolic enzyme or chemical agent, thereby releasing the mature functional protein in highly purified, highly active state.

REFERENCES:
patent: 4366246 (1982-12-01), Riggs
patent: 4468464 (1984-08-01), Cohen et al.
patent: 4503142 (1985-03-01), Berman et al.
patent: 4559302 (1985-12-01), Ingolia
patent: 4563424 (1986-01-01), Riggs
patent: 4571421 (1986-02-01), Itakura

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