Abstract

The aim of this study was conducted to investigate a method to production purify IgY and compare the capacity between Taiwan native Tsaiya ducks (layer type) and Leghorn laying hens. Ten Taiwan native Tsaiya ducks and 10 white Leghorn laying hens (20 wk-old) were injected with 250 ?L of complete Freund’s adjuvant containing bovine serum albumin (BSA, 200mg) during week 1 of the trial. The boost injection, during week 2 and 4 of the trial was comprised 250 ?L of incomplete Freund’s adjuvant containing BSA was conducted to promote IgY production. Eggs were collected for IgY purification from the week 5 of the trial. Yolk protein was first extracted using isopropanol or triton and followed by salting out with ammonium sulfate. After dialysis, gel filtration chromatography and affinity chromatography were used to purify the IgY antibody. Yolk proteins extracted by isopropanol or triton from duck eggs were 43% and 13%, and 61% and 16% from hen eggs, respectively. After salting out, the duck IgY had a purity of 53-57% with a titer of 26.2-29.9 unit/mg protein, while the hen IgY had a purity of 45-50% with 24-29.5 unit/mg protein. After purification by gel filtration chromatography, the purity of the duck IgY was increased to 89-90% with 44.24-50.62 folds purification and 415-451 unit/mg protein. The purity of the chicken hen IgY was increased to 85-87% with 29.85-30.38 folds purification and 240-277.6 unit/mg protein. After further purification by affinity chromatography, duck IgY purity was increased to 91-92% with 45-56.26 folds purification and 459-461 unit/mg protein; chicken hen IgY purity was increased up to 93-95% with 33.58-59.49 folds purification and 312-470 unit/mg protein. This procedure is promising for purification and production of the IgY antibody from Taiwan native Tsaiya ducks and/or white Leghorn laying hens, and both of them are good producer of IgY.

 

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