TY - JOUR T1 - Simple Methods for Purification of Beta-Lactoglobulin from Cow’s Milk while Preserving Its Antigenicity TT - تخلیص بتالاکتوگلوبولین از شیر گاو و مطالعه آنتی ژنیسیته آن JF - umsha JO - umsha VL - 9 IS - 1 UR - http://sjh.umsha.ac.ir/article-1-751-en.html Y1 - 2002 SP - 0 EP - 0 KW - Antigenicity / Beta-Lactoglobulin / Cow’s Milk Protein / Protein purification N2 - Beta-lactoglobulin (BLG) is a major whey protein in the milk of ruminants, e.g. cows, and such non-ruminants as horses. This protein is resistant under various conditions of pH and temperature. There is some evidence of absorption of some intact BLG in humans. Infants being fed with cow’s milk- based formulas are especially prone to develop adverse reactions against proteins, including BLG, absorbed intact through their intestinal wall that is more permeable than that of adults. Regarding its importance, purification of BLG for immunological and serological studies and for setting up such diagnostic tests as ELISA and RAST is undoubtedly necessary. The aim of this study was to introduce a simple, reproducible, fast and less expensive method for BLG purification while saving its antigenicity, as well. Whey (lactoserum) was prepared by isolating casein from defatted milk using HCl. Globulins were then precipitated in whey by half-saturated ammonium sulfate. The whey and precipitated globulins were used for Sephadex G-50 gel filtration and diethyl aminoethyl cellulose anion exchange chromatography. Using enzyme-linked immunosorbent assay (ELISA), Western blotting and ELISA inhibition assay, antigenicity of the purified protein was then evaluated. In gel filtration, BLG was eluted with the second peak. In anion exchange chromatography of ammonium sulfate precipitated proteins and of whey proteins, it was eluted with the second peak. Our results showed high purity and well-preserved antigenicity of BLG thus purified. M3 ER -