Scale-up of native beta-lactoglobulin affinity separation process.
Affinity separation of beta-lactoglobulin in its native form with all-trans-retinal immobilized on calcium bio-silicate was scaled up and applied to separate it from industrial sweet whey. Three different methods of mixing the modified calcium bio-silicate and whey for the interaction between all-trans-retinal and beta-lactoglobulin were tried at pilot scale. The three methods used were 1) a column packed with calcium bio-silicate, 2) a stirred tank, and 3) a fluidized bed column of calcium bio-silicate particles. Adsorption and desorption of beta-lactoglobulin were carried out at pH 5.1 and 7.0, using 0.01 and 0.1 M phosphate buffers, respectively. The phosphate buffer containing desorbed beta-lactoglobulin was concentrated 20 times using ultrafiltration and then freeze-dried. The packed column, stirred tank, and fluidized bed column produced beta-lactoglobulin with purity of 80, >95, and >95%, and recovery of 0.65, 2.88, and 2.88 g per kilogram of calcium bio-silicate, respectively. The comparative poor purity and recovery of beta-lactoglobulin in the case of the packed column was attributed to insufficient contact between the passing fluids and the calcium bio-silicate during adsorption, desorption, and intermittent washing. The fluidized bed column method, with a gentle mixing action, was considered the best suited for further scale up to the industrial level.
Vyas
H K
HK
Dairy Products Technology Center, California Polytechnic State University, San Luis Obispo 93407, USA.
Izco
J M
JM
Jiménez-Flores
R
R
D000327:Adsorption; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods*; D005374:Filtration; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins / Q000302:isolation & purification*; D008894:Milk Proteins / Q000737:chemistry*; D000067816:Whey Proteins
D007782:Lactoglobulins; D008894:Milk Proteins; D000067816:Whey Proteins