Isolation, Purification and Characterization of Ovalbumin from Hen Egg White and to Develop and isolate antibodies against it

Ovalbumin (54%) and lysozymes (3.5%) are among the major proteins in egg white that have many functional properties. Ovalbumin is a phosphoglycoprotein with a molecular size of 45 kDa and an iso-electric point of 4.5. Although, the functions of ovalbumin other than nutritional value for humans are not well known, it is widely used in cell culture, and has high potentials to be used as a drug carrier or to produce various functional peptidesThe aim was to isolate ovalbumin from hen egg white and to develop antibodies against it. First, egg white was homogenized and then crude ovalbumin was precipitated by salt fractionation using 36% Na2SO4. The protein was then subjected to dialysis and later filtered through Sephadex G-100 gel. Its molecular weight was estimated to be 35 kDa using SDS-PAGE. This purified ovalbumin was then injected into Wistar rats every 7 days over 4 weeks.Then blood was collected from these rats and centrifuged at 2000 rpm for 5 minutes. Serum was collected and its OD at 280 nm was measured to determine the IgG concentration which came out to be 10.13 mg/mL. Ouchterlony double immuno-diffusion assay between purified ovalbumin and commercially available ovalbumin showed identity pattern.