Figure 3
From: Development of novel detection system for sweet potato leaf curl virus using recombinant scFv
(A) The bio-panning result with sweet potato leaf curl virus (SPLCV) displayed on the yeast cells as an antigen. Sixty randomly selected colonies were measured after each panning round. The collected phages were bound to the target antigen and quantitatively confirmed using HRP-conjugated anti-M13 antibodies at an OD of 450 nm. Data are presented as means ± SEMs. (B) PCR detection of SPLCV in different tissues (phloem tissues and leaves) of sweet potatoes. Lane N is a no-template control. Lane H is virus-free sweet potato samples. Lane P1 and P2 are amplified with phloem tissue of SPLCV-infected samples, and 1, 2 are amplified viral DNA from sweet potato leaf samples. (C) Binding of scFv antibodies, determined by ELISA against SPLCV-infected plant leaves. SPLCV-infected sweet potato leaves were coated onto 96-well microtiter plates. Each scFv was detected using HRP-conjugated anti-M13 antibodies and TMB substrate solution. ELISA readings (OD450) were collected after 30 min of incubation in a TMB substrate at 25 °C. Clone numbers 10 and 12 are the selected scFv used for expression.
