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Fig. 5 | BMC Immunology

Fig. 5

From: Specific humoral response in cancer patients treated with a VEGF-specific active immunotherapy procedure within a compassionate use program

Fig. 5

Binding experiments using as ligands human VEGF-C, human VEGF-D and human VEGF. a Binding to VEGF-C or VEGF of different types of samples with specificity for VEGF. Myc-tagged proteins, VEGF-C or VEGF obtained from CHO cells (hVEGF-C CHO or hVEGF CHO, respectively), were captured using coated wells with a monoclonal antibody specific to myc-tagged proteins. Test samples were added and the binding of IgG or VEGFR1-Fcγ or VEGFR2- Fcγ or VEGFR3-Fcγ was detected with HRP-conjugated goat anti-human IgG antibody. This assay used as test samples a human serum positive for VEGF-specific IgG antibodies (PCS) and a human serum negative for VEGF-specific IgG antibodies (NCS). Control assays were bevacizumab (Bev) and VEGFR1, VEGFR2 and VEGFR3. b Histidine-tagged proteins (hVEGF-C CHO, hVEGF CHO, or commercially available VEGF-C and VEGF-D) were captured using nickel coated multiwell plates. Test samples were added and the binding of IgG or VEGFR1-Fcγ or VEGFR2- Fcγ or VEGFR3-Fcγ was detected with HRP-conjugated goat anti-human IgG antibody. This assay used as test samples a human IgG purified from a pool of serum belonging to patients classified with positive VEGF-specific IgG antibodies (IgG comp) and a human IgG isolated from pooled normal human serum (IgG neg). This assay used as control VEGFR3. Column bars represent the means of three replicates and error bars represent the standard deviations. p-values were calculated according to unpaired t-test. Statistical significance was considered as p < 0.05. ns non-significant

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