Figure 2

Purification of gp120_CN54⁺-Fc and OD_CN54⁺-Fc and reaction with MAbs b12 and 2G12. A. Protein present in the supernatant of recombinant baculovirus infected insect cells was purified by lectin and protein A chromatography and analysed by SDS-PAGE (tracks 1 and 2) and western blot (tracks 3 and 4) using an anti-human Ig conjugate. Tracks 1 and 3 – gp120_CN54⁺-Fc;Tracks 2 and 4 – OD_CN54⁺-Fc. Protein size markers indicated to the left are in kilodaltons (kDa). B. Purified protein was coated onto plastic at 10 μg/ml and used to assess the binding of monoclonal antibodies b12 or 2G12 as shown. The samples were (▲) – gp120_CN54⁺-Fc; (■) – OD_CN54⁺-Fc; (◆) – gp120_CN54⁺. Binding was detected by an anti-human Ig light chain conjugate.