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Blood, Vol. 113, Issue 16, 3716-3725, April 16, 2009
Specificity and affinity of human Fc receptors and their polymorphic variants for human IgG subclasses
Blood Bruhns et al.
113: 3716
Supplemental materials for: Bruhns et al
Files in this Data Supplement:
- Table S1. KA values (×105 M−1) of human Fcγ receptors for human IgG (PDF, 45.4 KB)
- Figure S1. Schematic representation of the extracellular domains of hFcγRIIA, hFcγRIIIA, and hFcγRIIIB (JPG, 48.4 KB)
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Three dimensional structures are accessible on the Protein Data Bank under accession numbers: 1H9V (hFcγRIIA), 1E4J (hFcγRIIIA) and 1FNL (hFcγRIIIB), http://www.rcsb.org/pdb. Only the amino acids corresponding to polymorphisms of hFcγRs are represented on the backbone of each molecule, using Discovery Studio 1.7.1, Accelrys Software. In hFcγRIIIB, NA1 amino acids are represented in green and black, NA2 amino acids in orange and black, and SH amino acids in orange and pink.
- Figure S2. FcRγ-chain–independent expression of FcγRIIIAV158 does not influence the binding of IgG1 and IgG3 (JPG, 60.9 KB)
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Histograms show the binding of anti-FLAG mAb (─) or its isotype control (▬), and the binding of monomeric IgG, heat-aggregated IgG or IgG-F′2 IC at indicated concentrations to FLAG-tagged hFcγRs on CHO transfectants. Concentrations of human IgG and PE-F(ab′)2 anti-human F(ab′)2 are indicated in this order. Solid grey histograms represent the binding of PE-F(ab′)2 anti-human F(ab′)2 alone. Two independent experiments gave similar results.
- Figure S3. Binding specificity of hFcγRs for monoclonal IgG (JPG, 152 KB)
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(A) Histograms show the binding of anti-FLAG mAb (─) or its isotype control (▬), and the binding of immune complexes made of biotinylated NIP12-BSA and anti-NIP mAbs subclasses to FLAG-tagged hFcγRs on CHO transfectants. Concentrations of human IgG and NIP12-BSA are indicated. (B) Binding specificity of low-dose therapeutic monoclonal IgG1-F′2 IC to hFcγRs. Histograms show the binding of anti-FLAG mAb (─) or its isotype control (▬), and the binding of indicated monoclonal IgG1-F′2 IC to FLAG-tagged hFcγRs on CHO transfectants. Concentrations of monoclonal IgG and PE-F(ab′)2 anti-human Ig are indicated. Monoclonal IgG1 used here were produced in CHO cells. Solid grey histograms represent the binding of PE-F(ab′)2 anti-human Ig alone. Two independent experiments gave similar results.
- Figure S4. Fitting of steady-state responses from data presented in Fig. 3A (JPG, 74.6 KB)
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The steady-state signal reached at the end of the injection, Req, was plotted against the concentration of IgG. Fitting using a Langmuir 1:1 model is represented for interactions of FcγRI, FcγRIIA and FcγRIIB/C for polyclonal IgG subclasses. Data correspond to the sensorgrams from the experiment presented in Fig. 3A.
- Figure S5. Fitting of steady-state responses from data presented in Fig. 3A (JPG, 58.9 KB)
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The steady-state signal reached at the end of the injection, Req, was plotted against the concentration of IgG. Fitting using a Langmuir 1:1 model is represented for interactions of FcγRIIIA and FcγRIIIB for polyclonal IgG subclasses. Data correspond to the sensorgrams from the experiment presented in Fig. 3A.
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