Full Length Research Paper

Isolation and antigenicity evaluation of β-lactoglobulin from buffalo milk

 X. Li^1,2, Z.L Luo^1,2, H.B Chen^1,2* and Y.S. Cao^1,2

¹State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, P.R. China.

²Jiangxi-OAI Joint Research Institute, Nanchang University, Nanchang 330047, P.R. China.

*Corresponding author. E-mail: chbgjy@hotmail.com.

Accepted 6 June, 2008

^{Buffalo β-lactoglobulin in phosphate buffer (0.02 M, pH6.8) was adsorbed on DEAE-Sepharose Fast Flow gel, and eluted with a linear gradient of NaCl (0-0.5 M) in 0.02 M phosphate buffer, pH 6.8. A further purification was performed on Sephadex G-75 gel by loading a concentrated and dialyzed fraction of samples containing buffalo β-lactoglobulin from ion-exchange chromatography, and seperating at a flow rate of 0.15 ml/min in 0.02 M phosphate buffer, pH 6.8. The purity of the isolated buffalo β-lactoglobulin was above 90% in comparison to the standard bovine β-lactoglobulin by SDS-PAGE and IEF-PAGE. The antigencity of the buffalo β-lactoglobulin was evualuted by indirect ELISA, Western-blotting and inhibition ELISA with anti-buffalo and anti-bovine β-lactoglobulin rabbit serum. The results showed that buffalo β-lactoglobulin could be seperated and purified by anion-exchange chromatography combined with gel filtration chromatography, and with a well-preserved antigenicity.}

^{Key words: Buffalo milk, milk allergy, β-lactoglobulin, antigenicity.}