6/20/2023 0 Comments Capto blue![]() In certain aspects, the disclosure herein relates to a method of reducing oxidation of tryptophan and/or methionine during purification in an albumin-fusion protein, the method comprising subjecting a composition comprising the albumin-fusion protein to the following purification processes: (a) an affinity matrix (b) an anion exchange matrix, wherein the albumin-fusion protein is eluted from the affinity matrix by applying an elution buffer comprising octanoate. The present invention relates to a method of purifying albumin-fusion proteins.Ĭitation or discussion of a reference herein shall not be construed as an admission that such is prior art to the present invention. For a platform approach to be successful, purification operations must be selective for the carrier protein. This is because in many cases the carrier protein makes up a large portion of the fusion protein, and thus there similar physiochemical characteristics between various fusion proteins. In addition to the benefits of half-life extension and ease of manufacturing, fusion proteins may also be able to take advantage of platform approaches to purification. All of the proteins are found in human plasma at high levels, mitigating the impact of increased levels due to the drug. Examples of fusion proteins include Fc-fusions, transferrin-fusions, and albumin-fusions. This option provides half-life extension similar to PEGylation however, it does not require the additional manufacturing steps (conjugation reaction and associated purification) since the fusion protein is expressed and purified as a single entity. In this case, the therapeutic protein is genetically fused to a second protein designed to extend half-life in vivo. Despite these challenges, PEGylation technology has been successfully employed in several commercial biopharmaceutical drugs.Ī second option for extending half-life is fusion protein technology. The disadvantage of PEGylation is that it requires a conjugation reaction step and often an additional purification step to remove unreacted PEG chains. In additional to half-life extension, PEGylation may also reduce immunogenicity, likely due to shielding of the protein surface by the inert PEG chains(s). ![]() One option to extended half-life is through PEGylation, a process by which poly(ethylene glycol), or PEG, is covalently attached to a protein through a number of available chemistries. To overcome this challenge, proteins and peptides can be conjugated or fused with other molecules. One disadvantage of protein drugs is they tend to have a short half-life in vivo. The use of proteins as potential therapeutic drugs has seen increased interest in recent years. The invention relates to the methods of purifying the abumin-fusion proteins, the purified proteins obtained from the method, and compositions comprising the purified albumin-fusion protein. The albumin-fusion protein may include scaffolds, such as those derived from the third fibronectin type III domain of human Tenascin C useful, for example. ![]() ![]() The low levels of oxidation of these residues allow the purified albumin-fusion protein to retain its relative potency and bioactivity. The present invention relates in general to a method of purifying albumin-fusion proteins having low levels of oxidation of the tryptophan and/or methionine residues of the protein. BACKGROUND OF THE INVENTION Field of the Invention 12, 2015 and having a size of 228 kilobytes. This application incorporates by reference a Sequence Listing submitted with the application via EFS-Web as a text file entitled “CD40L-300P1_SL.TXT” created on Mar.
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