Use of tris(hydroxymethyl) aminomethane for the stabilization of peptides, polypeptides and proteins

A method for reducing gelation/fibrillation/aggregation of peptides, polypeptides and proteins during up-/down-stream processing and purification, comprising the sequential steps:

• (a) adding an amount of tris(hydroxymethyl)aminomethane to an aqueous solution of peptide, polypeptide or protein sufficient to induce stabilization of the peptide, polypeptide or protein, wherein the tris(hydroxymethyl)-aminomethane is a buffering agent or an additive and wherein the aqueous solution further comprises an organic modifier which is selected from:
• methanol, ethanol, 1-propanol, 2-propanol, butanol, hexylene glycol, acetonitrile, and N-methyl-2-pyrrolidone;
• wherein the organic modifier is present in a concentration from about 1% w/w to about 70% w/w;
• (b) fractionating the aqueous solution of (a) by ion-exchange chromatography;
• and
• (c) fractioning the aqueous solution of (b) by reversed phase high performance liquid chromatography, or
• (b) fractioning the aqueous solution of (a) by reversed phase high performance liquid chromatography;
• and
• (c) fractionating the aqueous solution of (b) by ion-exchange chromatography, wherein the fractionating the aqueous solution by ion-exchange chromatography and fractioning the aqueous solution by reversed phase high performance liquid chromatography steps are performed without increasing back pressure on a chromatographic column, wherein gelation/fibrillation/aggregation of peptides, polypeptides and proteins during up-/down-stream processing and purification is reduced.