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Electronic Journal of Biotechnology ISSN: 0717-3458
  © 2010 by Universidad Católica de Valparaíso -- Chile
Vol. 13 No. 1, Issue of January 15, 2010
 


Figure 1. Quantitative receptor binding assay.
(a) Typical dose response curve for the quantitative potency assay. The pVGI.1(VEGF2) reference standard was pre-diluted to the starting concentration of 25 mg/mL before the 1:1 serial dilutions with the formulation buffer. The VEGF-2 in the conditioned media of the transfectants was then assayed for VEGFR3 binding activities (A450). The average of triplicate raw OD results was then plotted against the input DNA concentration in the transfection mix. The standard curve was modeled using a 4 parameter fit by SoftMax Pro v.3.1.
(b) The expected potency value vs. the measured potency value for the concentration range of 60-140% with the regression line (y = 0.949 x + 1.1617).
(c) Specificity of the binding reaction. The receptor binding detection was specific to the VEGF-C protein. Serially diluted protein was loaded on the plate and the binding signals were detected by OD450. The background was assessed using BSA buffer that did not contain any protein from the VEGF family.


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