Epigallocatechin gallate (EGCG) and β-lactoglobulin (βLg) were conjugated by covalent bonds to form EGCG-βLg conjugates. This conjugation causes structural and bioactivity changes in βLg, which in turn can be used as a possible approach for desensitization to allergens. In this study, the desensitization mechanism was investigated by monitoring βLg secondary structure and immunoglobulin E (IgE) combining capacity changes on the basis of the conjugation mechanism. Furthermore, the desensitization efficiency in vivo was evaluated through animal experiments. The results show that temperature influenced the conjugation by decreasing the binding affinities (Ka) and binding numbers (n) of EGCG. The conjugation of EGCG decreased βLg's IgE combining capacity by decreasing the β-sheet component and imparted antioxidant properties by the introduction of hydroxyl groups. In addition, animal experiment results indicated that βLg induced significant changes in the levels of IgE and inflammatory cytokines, and the relative abundance of small intestinal flora, linked to the inflammatory lesions and anaphylaxis symptoms. EGCG-βLg conjugates can suppress the allergic response, attenuating serum IgE and relieving the anaphylaxis symptoms.