The ELISA was carried out as described [13] (link). Briefly, a 96-well ELISA plate (Greiner Bio One, Germany) was coated overnight with 400 ng of purified HPV16 L1 VLPs per well and blocked with 5% skim milk in PBS containing 0.05% Tween 20 (PBST). The plate was incubated with 250 ng/mL of anti-HPV16 neutralizing monoclonal antibodies (Mabs), H16.V5 or H16.E70, for 1 h at 37°C. The Mabs bound to VLPs were detected using HRP-conjugated goat anti-mouse IgG antibody (Bethyl Laboratories, USA). Color reactions were developed with o-phenylenediamine (Sigma, USA), and optical density was measured at an absorbance of 492 nm.
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