Capture of lactoferrin and lactoperoxidase from raw whole milk by cation exchange chromatography

Abstract

The production of high-value dairy proteins such as lactoferrin and lactoperoxidase normally requires extensive pre-treatments of milk to remove fat and caseins by centrifugation, precipitation, Ca 2+ chelation and/or filtration. Similarly, fat and caseins are normally removed prior to capture of recombinant proteins from the milk of transgenic animals. Such pre-treatments can result in significant loss of protein yield and/or activity. In this paper we demonstrate that it is possible to pass significant quantities of raw, untreated milk through a 5 cm high chromatography column packed with SP Sepharose Big Beads™ (GE Healthcare, Uppsala, Sweden) without exceeding the maximum allowable backpressure, provided that the processing temperature is kept nominally around milking temperature (35 to 37 o C).Results show that more than 100 column volumes of raw milk could be loaded at 300 cm/hr before breakthrough of lactoperoxidase occurred. The dynamic capacity for adsorbing lactoferrin and lactoperoxidase simultaneously under these conditions was approximately 48.6 mg/mL of resin. Minor leakage (4.6% of the feed concentration) of lactoferrin occurred throughout the loading process but major breakthrough occurred only after approximately 100 column volumes was loaded.

Fractionation of β‐Lactoglobulin from whey by mixed matrix membrane ion exchange chromatography

“…Figure 2 shows the binding capacity of intact MP500 resin for b-Lac at different pH values. The maximum binding capacity was achieved around pH 5-6 which is near the pI of b-Lac (pH 5.35-5.49 (Fee and Chand, 2006)). Intuitively, one would expect the binding to increase as the pH of binding deviates upwards from the pI of the protein, as the protein would be more highly charged at higher pH values.…”

Processing Foods Free from Dairy Proteins

“…Selective precipitation, large pore ultrafi ltration, and ion exchange chromatography are used to isolate the major proteins β -lactoglobulin ( β -Lg), α -lactalbumin ( α -la), and glycomacropeptide from cheese whey [10] . Specialized techniques such as cation exchange chromatography are used for the purifi cation of minor proteins such as lactoferrin (LF) and lactoperoxidase [31] . Additionally, enzymatic whey protein hydrolysates are produced from whey or the individual protein isolates for use in food applications.…”

Optimization of Lactoperoxidase and Lactoferrin Separation on an Ion-Exchange Chromatography Step

“…Whey proteins have been adequately separated into different fractions, the major and minor proteins [4][5][6][7][8]. Isolation and purification of major proteins, e.g., β-lactoglobulin, α-Lactalbumin and serum albumin, through various chromatographic and membrane-separation techniques have been extensively studied [7,9,10].…”