title: Characterization of cryogel monoliths for extraction of minor proteins from milk by cation exchange.
abstract: Extraction and purification of high-value minor proteins directly from milk without pre-treatment is a challenge for the dairy industry. Pre-treatment of milk before extraction of proteins by conventional packed-bed chromatography is usually necessary to prevent column blockage but it requires several steps that result in significant loss of yield and activity for many minor proteins. In this paper, we demonstrate that it is possible to pass 40-50 column volumes of various milk samples (raw whole milk, homogenized milk, skim milk and acid whey) through a 5 mL cryogel chromatographic column at 550 cm/h without exceeding its pressure limits if the processing temperature is maintained above 35 degrees C. The dynamic binding capacity obtained for the cryogel matrix (2.1 mg/mL) was similar to that of the binding capacity (2.01 mg/mL) at equilibrium with 0.1 mg/mL of lactoferrin in the feed samples. The cryogel column selectively binds lactoferrin and lactoperoxidase with only minor leakage in flowthrough fractions. Lactoferrin was recovered from elution fractions with a yield of over 85% and a purity of more than 90%. These results, together with the ease of manufacture, low cost and versatile surface chemistry of cryogels suggest that they may be a good alternative to packed-bed chromatography for direct capture of proteins from milk.
authors.affiliation: Department of Chemical and Process Engineering, University of Canterbury, Private Bag 4800, Christchurch 8140, New Zealand.
authors.forename: Jagan M
authors.initials: JM
authors.lastname: Billakanti
authors.forename: Conan J
authors.initials: CJ
authors.lastname: Fee
mesh_terms: D000818:Animals; D001798:Blood Proteins; D002411:Cation Exchange Resins; D002852:Chromatography, Ion Exchange / Q000379:methods*; D059985:Cryogels; D005353:Fibronectins; D020100:Hydrogels; D008892:Milk / Q000737:chemistry*; D008894:Milk Proteins / Q000302:isolation & purification*
chemical_list: D001798:Blood Proteins; D002411:Cation Exchange Resins; D059985:Cryogels; D005353:Fibronectins; D020100:Hydrogels; D008894:Milk Proteins