Nanoparticle orientationally displayed antigen epitopes improve neutralizing antibody level in a model of porcine circovirus type 2

Ding, Peiyang; Zhang, Teng; Li, Yafei; Teng, Man; Sun, Yaning; Liu, Xiao; Chai, Shujun; Zhou, Enmin; Jin, Qianyue; Zhang, Gaiping

Detection of anti-cap, anti-cap (169-180), and anti-PcV2 antibodies Antibody titers were measured by ELISA. Briefly, Cap protein or BSA-Cap (169-180) was diluted to 5 μL/mL by carbonate buffer (pH 9.6), then coated on 96-well microtiter plates, and incubated at 4°C overnight. For detection of antibodies specific to PCV2, a commercial PCV2 antibody test kit (BioChek, Reeuwijk, Holland) was used. The ELISA microtiter plates were coated with purified and inactivated PCV2. After blocking with skim milk (2.5% in PBST), appropriate dilutions of serum samples were added to the wells and the plates were then incubated at 37°C for 1 h. After five washes with PBST, HRP-conjugated goat anti-mouse IgG and IgG subtype were added for 1 h at 37°C. The reaction was developed using TMB. The OD value of each well was measured at 450 nm using an ELISA microplate reader.