Preparation and activity research of chicken yolk antibody targeting SARS-CoV-2
Objective:To prepare highly effective anti-SARS-CoV-2 yolk antibody(IgY)using IgY technology and detect its immunological activity.Methods:Laying hens were immunized by the inacti-vated SARS-CoV-2 vaccine,and IgY was extracted by freeze-thawing water dilution,ammonium sul-fate precipitation,and dialysis for concentration and purification,and the IgY extraction method was optimized.The extraction and purification of IgY were characterized by SDS-PAGE,the concentra-tion of the purified proteins was measured by BCA,and the potency of the antibodies against recombi-nant SARS-CoV-2 S1 and N proteins,as well as their thermal stability,were measured by indirect ELISA.Balb/c mice were given 15 days of a conventional dose of IgY and 30 days of a large dose of IgY through oral and nasal drops,respectively.To confirm the safe dosage range of IgY,ELISA was used to determine the immune reactivity of mice serum to IgY and the expression of IL-1β,IL-6,IFN-γ,and TNF-α in mice.Results:Purificated IgY SDS-PAGE results indicated that the modified extraction and purification of IgY is feasible.The titer of purified IgY reached the highest in the 2nd week after immunization.When the concentration of IgY is 200 μg/mL,the titer of IgY against re-combinant SARS-CoV-2 S1 protein and N protein reaches 1∶256.The activity of IgY remains stable after treating in water for at 25-60℃4 hours.The conventional dose of IgY can not induce a signifi-cant immune response in mice.During the long-term high dose of IgY by oral and nasal administra-tion,20 mg/mL IgY treatment significantly increased the secretion of IL-1β,IL-6,IFN-γ,and TNF-α in the serum of treated mice.Conclusion:IgY technology has the potential to rapidly obtain highly efficient IgY against different antigens,and the IgY produced by it has good thermal stability and bio-safety,but the dose of IgY should be avoided to be more than 50 mg/kg for long-term use so as not to aggravate the burden of kidney function.
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