Delta Biotechnology Limited

Novozymes Biopharma UK Limited

Novozymes Delta, Limited

Alan Victor Quirk

Richard A Johnson

John Rodney Woodrow

Stephen Berezenko

Patricia Carol Wood

Andrew Robert Goodey

Hendrik Van Urk

Stephen James Burton

Darrell Sleep

Mark J Wilson

Steven James Burton

Darell Sleep

David St Coghlan

Patricia Carolyn Wood

A process is provided for the preparation of albumin from a yeast culture medium which has extremely low levels of or is essentially free of colorants, metal ions, human proteins, host proteins, fragments of albumin, polymers or aggregates of albumin and viruses, and which is essentially non-glycated, relatively high in free thiol and with an intact C-terminus. The process comprises separating the yeast from the culture medium to yield an albumin solution; subjecting the albumin solution to positive mode cation exchange, affinity, and positive mode anion exchange chromatography; and concentrating and formulating the albumin with sodium octanoate and sodium chloride.

Process of high purity albumin production

A process is provided for the preparation of albumin which has extremely low levels of or is essentially free of colorants, metal ions, human proteins, host proteins, fragments of albumin, polymers or aggregates of albumin and viruses, and which is essentially non-glycated, relatively high in free thiol and with an intact C-terminus. The process comprises passing albumin (preferably expressed and secreted by transformed yeast) through positive mode cation exchange and then positive mode anion exchange chromatography. Other steps may also be employed, ultrafiltration, gel permeation chromatography, affinity chromatography binding the albumin by using blue dyes) and affinity chromatography binding contaminants by using an aminophenylboronic acid resin. Elution of albumin, with a compound having affinity for albumin, from a material having no specific affinity for albumin is also disclosed, as is removal of ammonium ions with a counter-ion.

The present invention relates to a process for purifying the protein human serum albumin extracted from serum or recombinant human albumin produced by transforming a microorganism with a nucleotide coding sequence encoding the amino acid sequence of human serum albumin. The present invention provides highly purified albumin.

A process is provided for the preparation of albumin which has extremely low levels of or is essentially free of colorants, metal ions, human proteins, fragments of albumin, polymers or aggregates of albumin, and viruses, and which is relatively non-glycated, relatively high in free thiol and with an intact C-terminus. The process comprises passing albumin (preferably expressed and secreted by transformed yeast) through two chromatography purifications, ultrafiltering the product, passing through two further chromatography steps and again ultrafiltering the product.

High purity albumin

When proteins are purified using a protein-binding dye immobilized on a chromatographic matrix, the dye or a portion/derivative may leak into the eluant. An ion-exchange resin (e.g. Dowex-1) and a disrupting material (e.g. salt and a fatty acid such as sodium octanoate) are used to separate the dye from the protein to overcome the protein of the leaking dye.

Purification of proteins

Yeast strains

A process for the preparation of albumin which has extremely low levels of or is essentially free of colorants, metal ions, human proteins, fragments of albumin, polymers or aggregates of albumin, and viruses, and which is relatively non-glycated, relatively high in free thiol and with an intact C-terminus. The process comprises passing albumin (preferably expressed and secreted by transformed yeast) through two chromatography purifications, ultrafiltering the product, passing through two further chromatography steps and again ultrafiltering the product.

High purity albumin and method of producing

Problem: when proteins are purified using a protein-binding dye immobilised on a chromatographic matrix, the dye or a portion/derivative may leak into the eluant. Solution: an ion-exchange resin (e.g. Dowex-1) and a disrupting material (e.g. salt and a fatty acid such as sodium octanoate) are used to separate the dye from the protein.

Growing community of inventors

Loughborough, United Kingdom

Alan Victor Quirk