Application of peptide chromatography for the isolation of antibodies from bovine skim milk, acid whey and colostrum

• HWRGWV peptide resin showed an equilibrium adsorption capacity of 23 ± 0.58 mg mL−1 of resin.
• HWRGWV peptide resin showed interactions to both bovine IgG and IgA antibodies.
• Bound antibodies were recovered at mild elution conditions of pH 4.0.
• Recovered antibodies showed >85% purity for colostrum.
• HWRGWV resin may be suitable for the purification of various antibodies from milk.

Protein A mimetic peptide ligands have several benefits over conventional Protein A/G ligands, namely that they are small in size, have low production costs, are stable over a wide range of pH values and can withstand cleaning by harsh sanitization agents such as sodium hydroxide. In this paper, a hexamer peptide (HWRGWV) affinity matrix was used for the isolation of bovine immunoglobulins from various dairy streams (skim milk, acid whey and colostrum). Bound immunoglobulins were recovered in elution buffer (0.2 M sodium acetate buffer, pH 4.0) fractions with a purity of >85% in a single step. The peptide resin has achieved a maximum equilibrium adsorption capacity of 23 ± 0.58 mg mL−1 of resin for bovine IgG and had a dynamic binding capacity of 11.8 ± 0.03 mg mL−1 at residence time of 2 min. These results suggest that the hexamer peptide chromatography could potentially be used for the selective purification of bovine immunoglobulins from dairy streams. This method has promise as an alternative to conventional Protein A/G chromatography for direct capture of immunoglobulins from streams containing relatively high immunoglobulin concentrations such as colostrum, transgenic or hyper-immune milk.