Process for making antifusogenic fusion peptides that form inclusion bodies
A process is provided for producing an antifusogenic peptide by producing a fusion peptide of from about 14 amino acids up to 70 amino acids in a prokaryotic host cell under conditions in which inclusion bodies are formed. The antifusogenic peptide recovered from the inclusion bodies is a fragment cleaved from the fusion peptide which comprises an antifusogenic peptide.
1. A process for the production of an antifusogenic peptide by producing a fusion peptide of a length of from 14 amino acids up to 70 amino acids in a prokaryotic host cell, comprising the steps, under such conditions that inclusion bodies of said fusion peptide are formed, of:
(a) expressing in said host cell a nucleic acid encoding said fusion peptide consisting of a first peptide which is an antifusogenic peptide of a length of 10 to 50 amino acids and a second peptide of a length of 4 to 30 amino acids, said first peptide being N-terminally linked to said second peptide;
(b) cultivating said host cell to produce said infusion bodies and;
(c) recovering said antifusogenic peptide from said inclusion bodies, wherein said recovered antifusogenic peptide consists of said fusion peptide or a peptide comprising the antifusogenic peptide of 10 to 50 amino acids resulting from cleavage of said fusion peptide.
2. The process according to claim 1 , wherein the second peptide consists of a peptide selected from:
(a) peptides which consist of from 4 amino acids to 20 amino acids,
(b) cleavable peptide linkers which consist of from 4 amino acids to 10 amino acids, and
(c) peptides consisting of
(i) a peptide consisting of up to 20 amino acids, and
(ii) a cleavable peptidic linker which consists of up to 10 amino acids, wherein the cleavable peptide linker is located between the N-terminus of the first peptide and the C-terminus of the peptide according to (i).
3. The process according to claim 1 , wherein the ratio of the molecular weight of the first peptide to the molecular weight of the second peptide is from 10:1 to 1:2.
4. The process according to claim 1 , further comprising recovering the inclusion bodies produced in accordance with step (a) from said host cells, and solubilizing the recovered inclusion bodies, to recover said antifusogenic peptide from the inclusion bodies.
5. The process according to claim 4 , wherein the first peptide is cleaved from the second peptide of the fusion peptide during or after solubilization of the recovered inclusion bodies.