Volume 9, Number 1 (Scientific Journal of Hamadan University of Medical Sciences-Spring 2002)                   Sci J Hamadan Univ Med Sci 2002, 9(1): 0-0 | Back to browse issues page


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Neyestani T. Simple Methods for Purification of Beta-Lactoglobulin from Cow’s Milk while Preserving Its Antigenicity. Sci J Hamadan Univ Med Sci . 2002; 9 (1)
URL: http://sjh.umsha.ac.ir/article-1-751-en.html

Abstract:   (630 Views)

          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.

     
Type of Study: Original | Subject: Special

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