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. 2018 Jun;16(1):99-103.
doi: 10.1016/j.jgeb.2017.10.003. Epub 2017 Oct 10.

Production and purification of IgY antibodies from chicken egg yolk

Wala Ahmad Amro  1 , Wael Al-Qaisi  2 , Fawzi Al-Razem  1
Affiliations

Production and purification of IgY antibodies from chicken egg yolk

Wala Ahmad Amro et al. J Genet Eng Biotechnol. 2018 Jun.

Abstract

The isolation of polyclonal antibodies from the serum of immunized mammals has significantly contributed to scientific research and diagnosis. The fact that recent technologies allow the production of antibodies in the yolk of eggs laid by hens, has led to the development of an alternative method for antibody generation that is less stressful to animals. As hens are kept under almost all their natural conditions, antibodies are isolated from the collected eggs; this technology is expected to become an interesting alternative to the conventionally serum-based techniques that eventually require to sacrifice the animal. Here we present a modified protocol for the isolation of IgY antibodies from immunized chickens and provide comparison between two chicken breeds in relative to IgY yield per egg. Our results show the possibility of generating large quantities of highly pure IgY from chicken eggs and also show large differences in the yield of IgY production between the two studied breeds. The results of this work indicate that IgY technology can be used for the production of primary antibodies for immunological work and disease diagnosis.

Keywords: Chicken egg yolk; Gel filtration chromatography; IgY; Salmonella typhimurium.

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Figures

Fig. 1
Fig. 1
A highly pure IgY from immunized Single Comb White Leghorns egg yolk after the third dose of vaccine was obtained using gel filtration chromatography. Y-axis represents the absorbance units (AU), X-axis represents the time (minutes). Pure IgY from fractions No. 14–17 were collected for further quality testing and with 0.13 AU for the two chains of the IgY.
Fig. 2
Fig. 2
The SDS-PAGE for the purity at each stage in purification and precipitation steps from egg yolk. Lane 1: PiNK Plus Prestained Protein Ladder, lane 2: the sample after 3.5% PEG precipitation, lane 3: the sample after 8.5% PEG precipitation, lane 4: the sample after 12% PEG precipitation. The IgY HC ≈ 65 kDa, and the LC ≈ 27 kDa.
Fig. 3
Fig. 3
The SDS-PAGE profile of IgY. The two IgY chains appeared on the SDS PAGE by using 10% resolving SDS-PAGE gel. The HC heavy chain with 65 kDa, LC the light chain with 27 kDa. lane 1 = BLUeye Prestained Protein Ladder, lane 2–7 = purified IgY, lane 8 = PiNK Plus Prestained Protein Ladder.
Fig. 4
Fig. 4
Identification of IgY by Western blot. Lane 1 = BLUeye Prestained Protein Ladder, lane 2–7 = purified IgY, lane 8 = PiNK Plus Prestained Protein Ladder. Result of Western blot test using Anti-chicken IgY (IgG) (whole molecule) alkaline phosphatase antibody produced in rabbit to detect the heavy chains and the light chains of chicken IgY. The LC: light chain ≈ 65 kDa, the HC: heavy chain ≈ 27 kDa.
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References

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