Journal: Virology Journal

Article Title: Transmission of human hepatitis C virus from patients in secondary cells for long term culture

doi: 10.1186/1743-422X-2-37

Figure Lengend Snippet: Dot blot analysis of HCV proteins binding to IgG from patient sera . HCV proteins were obtained from tissue culture fluid. Protein preparations were serially diluted (1, 10 -1 , 10 -2 , 10 -3 ) and were dot blotted onto a nitrocellulose membrane. These blots were then treated with patient antibodies. The figure shows (A) Reactions of patient IgG against dilutions of IgG depleted patient sera, CIMM-HIV cell culture supernatants from different cell lines (HCV infected B-cells, HCV infected human neuronal precursor T and M cells), or commercial antigens (NS4 and Core antigen), or uninfected B-cells. (B) Reactions of patient IgG against dilutions of various HCV isolates grown in vitro as described before. All HCV isolates were from the first transfer to fresh B cells (T1) except for isolates 314T2 (second transfer) and PCLBT4 (fourth transfer). These infected cells have been in culture for varying periods of time, including over three years for PCLBT4.

Article Snippet: For the dot-blot assay, 2 μl of various protein samples (undiluted to 10 -3 ) were diluted to 25 μl using TBS and were dot blotted onto a nitrocellulose membrane (0.22 μ, Micron Separations Inc. Westboro, MA).

Techniques: Dot Blot, Binding Assay, Membrane, Cell Culture, Infection, In Vitro