Immunochemical characterization of purified buffalo beta-lactoglobulin


  • Ranjit Aich College of Veterinary Science and Animal Husbandry, Mhow, Madhya Pradesh, India
  • Subhasis Batabyal West Bengal University of Animal and Fishery Sciences, Belgachia, Kolkata, India
  • Siddhartha Narayan Joardar West Bengal University of Animal and Fishery Sciences, Belgachia, Kolkata, India


Beta-lactoglobulin, Buffalo, Western blotting, Indirect ELISA, Dipstick ELISA


Beta-lactoglobulin (β-lg) is a major whey protein found in cow milk and other ruminants, deer, bison, and buffalo. β-lg is considered as one of the main allergenic components in bovine milk and therefore the modification of β-lg is considered as a promising treatment for milk allergy. Thus the present study was conducted to assess the antigenicity of purified β-lg that can be exploited for detection of buffalo milk-protein intolerance. β-lg was purified from milk of Murrah buffalo of Haringhata Farm, West Bengal (India) by combination of gel filtration (using Sephacryl S-200) and anion-exchange chromatography (using DEAE-Sepharose). The molecular weight was derived as 18.05 kDa in 15 percent one-dimensional SDS-PAGE. Antigenicity (seroreactivity) of purified β-lg was evaluated by double immuno diffusion (DID), western blotting, dipstick ELISA and indirect ELISA using hyperimmune sera raised in rabbit against purified buffalo β-lg. In indirect ELISA, titer of rabbit serum against buffalo β-lg was observed to be in between 200-400. Therefore, the study revealed that purified buffalo β-lg is highly antigenic and a seroreactive protein and may be used for detection of buffalo milk proteinintolerance using rapid, user-friendly, cost effective sero-diagnostic technique/tool


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How to Cite

Aich, R., Batabyal, S., & Joardar, S. N. (2016). Immunochemical characterization of purified buffalo beta-lactoglobulin. Buffalo Bulletin, 35(2), 269–275. Retrieved from



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