Figure 2

Detection of anti-HCV antibodies in HCV RNA and antibody positive human sera with recombinant HCV proteins. 1 μg of each baculovirus-expressed and preparative SDS-PAGE-purified recombinant HCV protein was loaded onto 10–20% Tris-glycine polyacrylamide gradient gels. Core, NS2, NS3, NS4A, NS4B, and NS5A were loaded on one gel, and E1, E2, and NS5B on another gel, respectively. 3 μg of Sf 9 cell extract was also loaded onto one gel as a control. Proteins separated on gels were transferred to nylon membranes, sliced and stained with serially diluted human serum obtained from HCV RNA and antibody positive patients. The following dilutions were used (lane 1) 1:100, (lane 2) 1:500, (lane 3) 1:2500, (lane 4) 1:12500 and 1:62500 (not shown). After incubation with secondary Abs, the bands were visualized by 3-amino-9-ethylcarbazole (AEC). A. An example of highly positive human serum number 36 is shown, B. an example of a weakly positive human serum number 17 is shown.