Systems and methods for producing collagen 7 compositions

A production system for producing a collagen 7 composition, comprising host cells modified to express human recombinant collagen 7 (rCol7) and/or functional variants thereof,

• wherein the host cells are modified to express:
• (a) human collagen 7 alpha chain polypeptide, and/or a functional variant thereof,
• (b) alpha polypeptide of prolyl 4-hydroxylase, or a functional variant thereof,
• (c) beta polypeptide of prolyl 4-hydroxylase, or a functional variant thereof, and
• (d) heat shock protein 47, or a functional variant thereof,
• and wherein the collagen 7 composition comprises human rCol7, functional variants of human collagen 7, or the combination thereof.

A modified host cell for producing a collagen 7 composition comprising human rCol7, and/or functional variants thereof, wherein the host cell is transformed to express:

• (a) human collagen 7 alpha chain polypeptide, or a functional variant thereof;
• (b) alpha polypeptide of prolyl 4-hydroxylase, or a functional variant thereof, wherein the alpha polypeptide of prolyl 4-hydroxylase is an alpha-1, an alpha-2, or an alpha-3 polypeptide;
• and
• (c) beta polypeptide of prolyl 4-hydroxylase, or a functional variant thereof.

A modified host cell for producing a collagen 7 composition comprising human rCol7, and/or functional variants thereof, wherein the host cell is transformed to express:

• (a) human collagen 7 alpha chain polypeptide, or a functional variant thereof;
• (b) alpha polypeptide of prolyl 4-hydroxylase, or a functional variant thereof, wherein the alpha polypeptide of prolyl 4-hydroxylase is an alpha-1, an alpha-2, or an alpha-3 polypeptide;
• (c) beta polypeptide of prolyl 4-hydroxylase, or a functional variant thereof;
• and
• (d) heat shock protein 47, or a functional variant thereof.

A method for producing a human collagen 7 composition comprising:

• (a) culturing host cells in a serum free medium, wherein the host cells are modified to express,
• (i) human collagen 7 alpha chain polypeptide, or a functional variant thereof;
• (ii) prolyl 4-hydroxylase, or a functional variant thereof, wherein the prolyl 4-hydroxylase comprises an alpha prolyl 4-hydroxylase polypeptide and a beta prolyl 4-hydroxylase polypeptide, and
• (iii) heat shock protein 47, or a functional variant thereof;
• and
• (b) collecting the culture media, and
• (c) purifying said human collagen 7 composition.