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The wells were blocked with DPBS (pH 7.4)/4% (w/v) skimmed milk powder (Sigma-Aldrich) (PBS/M) (250 L/well) for 1 h at room temperature and washed three times with PBS/0.05% (v/v) Tween 20 (PBS/T) at either pH 5.5 or pH 7.4

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Based on these data, our three-cassette construct, which allows simultaneous expression of tag-free VP0, VP3, and VP1, appears to be much more efficient than the other two strategies in terms of the final PV1 D-antigen yields achieved

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