Isolation and Estimation of Chicken Immunoglobulins (IgY) from Egg Yolk by Optimizing Polyethylene Glycol (PEG) Precipitation Method
Journal Title: Scholars Journal of Agriculture and Veterinary Sciences - Year 2017, Vol 4, Issue 7
Abstract
Immunoglobulin Y (IgY) is found in high concentrations in the egg yolk of chicken. Birds concentrate immunoglobulins into their egg yolks to protect the offspring. The practical application of IgY in research, diagnostics and functional food is limited due to complex and time-consuming purification procedures. The objective of this study was to develop an economic, simple, safe, large-scale separation method for IgY from egg yolk. In the present study, we determined egg yolk IgY concentration in six lines of chicken by optimizing polyethylene glycol (PEG) precipitation method. Egg yolk samples were collected from studied lines of chicken. PBS (Phosphate Buffer Saline) was taken twice of the egg yolk volume and mixed with the yolk. Thereafter 3.5% PEG (Polyethylene glycol) of the total volume was added and centrifuged (4°C) for 30 min at 4500 rpm. The supernatant was poured through a folded filter paper into a new tube and 8.5 % PEG was added to the tube, and centrifuged again. The supernatant was discarded and PBS was added to make a volume of 10 ml. The solution was mixed with 12 % PEG and centrifuged again. The pellet was carefully dissolved in 2 mL PBS. Finally, the isolated IgY samples were stored at -20°C until further processing. SDS-PAGE was performed to check the quality of IgY. The purity of IgY was determined by electrophoresis. The result showed that the yield of isolated IgY was varied between 206.015 and 392.030 mg per egg in the experimental lines of chicken. In this optimized method, the yield of IgY was the highest 30.904 ± 7.621 mg/mL of egg yolk and the lowest was 16.753 ± 5.282 mg/mL. This optimized precipitation method was simple with a higher yield of IgY, more efficient and useful for the large-scale preparation of IgY from egg yolk.
Authors and Affiliations
ChoudhuryNafisaBinte Hussain
Keywords
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