Extraction and Quantitation of IgY from Poultry Eggs

Wee, Mei Jee (2025) Extraction and Quantitation of IgY from Poultry Eggs. Final Year Project (Bachelor), Tunku Abdul Rahman University of Management and Technology.

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Abstract

Immunoglobulin Y (IgY) is an antibody that can be found in avian species. IgY presents an alternative prospect - in clinical and research applications - to the more commonly used mammalian-based antibodies due to its specificity, stability, and non-reactivity with the human immune system. Conventional methods of extracting IgY from blood serum are complicated and costly. The main objective of this study is to explore an optimal extraction method of IgY from poultry eggs. The study aims to establish a method that is cost-effective and environmentally sustainable without compromising IgY yield. Here, we show simple and low-cost IgY extraction from poultry eggs using water dilution and corn starch. A 1:4 yolk-to-distilled water ratio and cold centrifugation at 4°C were used to maintain the protein integrity. The purification process, which primarily used traditional kitchen salt, improved protein recovery. The samples containing IgY were run in SDS-PAGE, and the results of different extraction methods were compared. SDS-PAGE results were quantified using ImageJ software, calculated based on the integrated density by summing all the pixel values within the selected area of the protein bands. This value reflects the total band intensity and is used to estimate the amount of protein present. Based on SDS-PAGE results, the WD method consistently produced the total band intensity compared to corn starch at both concentrations. For chicken eggs, the WD method yielded an intensity of 2.10 × 10⁶ for the heavy chain and 1.36 × 10⁶ for the light chain. Quail eggs followed with 1.95 × 10⁶ (heavy chain) and 1.26 × 10⁶ (light chain). An additional experiment was done for duck eggs using a 1:10 dilution of delipidation agents, the WD method again showed the best results with intensity of 1.56 × 10⁶ (heavy chain), 2.09 × 10⁶ (truncated heavy chain), and 1.15 × 10⁶ (light chain). The optimized protocols achieved a 50% reduction in processing time, with the cost of salt per egg estimated at RM 0.012 for the WD method and RM 0.04 for the corn starch method, making the approach both time-efficient and cost-effective. Keywords: Immunoglobulin Y (IgY); Poultry eggs; Water-dilution (WD); Corn starch; SDS-PAGE