Separation of minor protein components from whey protein isolates by heparin affinity chromatography

Trends in whey protein fractionation

“…Konecny et al (1994) used thiophilic chromatography on a T-gel (thiophilic gel composed of divinylsulfone activated-agarose coupled with 2-mercaptoethanol) to purify IgG from sweet cheese whey and found this method suitable for large-scale whey IgG isolation. Ounis et al (2008) used heparin affinity chromatography to extract minor proteins from whey protein isolates. The results suggested that this method can be used to concentrate the minor cationic proteins such as LF and other growth factors.…”

In Vitro Digestion of Proteins and Growth Factors in a Bovine Whey Protein Extract as Determined Using a Computer-Controlled Dynamic Gastrointestinal System (TIM-1)

“…The reversed-phase high-performance liquid chromatography (RP-HPLC) method with standard curves as described by Ben Ounis et al [25] was used to quantify the major BWPE proteins in the samples. Briefly, a Resource™ RPC 1 ml column (Amersham Biosciences, Uppsala, Sweden) connected to an HPLC system from Waters (Milford, MA, USA) was operated at 25°C and a flow rate of 1 mL/min.…”

“…Quantification of TGF-β2 was done using the ELISA method of Ben Ounis et al [25], with the sample concentration adjusted to 50 μg/mL to place the TGF-β2 content in proper range. A TGF-β2 calibration curve (31.25-1,500 pg/mL) was prepared with purified porcine TGF-β2.…”

Isolation and Purification of Bioactive Proteins from Bovine Colostrum

“…Analogously, Chen made a preparation of Lysozyme bound sepharose 4 fast flow gel which was applied to isolate IgY in the affinity chromatrography. This research showed the binding capacity was lower and the dissociation constant was higher than both of the monoclonal antibody immunoaffinity column chromatography; in addition, this Lysozyme bound sepharose 4 immunoaffinity column was competent in sparating IgY specific against Lysozyme from yolk (Chen et al, 2002) 3.2.3 Novel affinity chromatography process for the purification of bioactive protein from Bovine colostrum Ounis once used heparin affinity chromatography to separate the protein components from two whey protein solutions which produced by ion-exchange chromatography (IEC-WPI) and microfiltration / ultrafiltration (MF/UF-WPI) respectively (Ounis, et al, 2008). After the column was equilibrated, WPI solution was passed through the column at a flow rate of 1 mL/min, then the column was washed by 0.01 M phosphate buffer , in the wake of this, sequential elution steps were executed with 0.01 M phosphate buffer containing 0.5, 1.0 or 2.0 M NaCl.…”