Table 1.
Antibodies, Neutralizing Agents, and Growth Factors/Cytokines Used in This Study
| Agent | Type/use | Source | Target | Effective dose∗ | Producer | Notes |
|---|---|---|---|---|---|---|
| Aflibercept | sR-Decoy | Engineered | VEGFs A-E, PlGF | 40 μg/μL (0.9 mg) | Regeneron | † |
| α-PDGF-AB | nAb | Human | PDGFs A, AB, B | Per manuf. | Millipore | |
| α-PDGF-C | nAb, quant-IB | Human | PDGF-C | 1:2000 serum Ab | ZymoGenetics (Seattle, WA) | 21 |
| α-PDGF-D | nAb, quant-IB | Human | PDGF-D | Per manuf. | R&D Systems (Minneapolis, MN) | |
| α-CTGF | nAb, quant-IB | Goat | CTGF (see Notes) | 200 ng/mL | Santa Cruz Biotechnology (Santa Cruz, CA) | 24 |
| sEGFR | sR-Decoy | Engineered | EGF, TGF-α | 4 μg/mL | R&D Systems | 27 |
| α-FGF-2 | nAb | Mouse | FGF-2 (bFGF) | Per manuf. | R&D Systems | |
| α-G-CSF | nAb | Rabbit | G-CSF | Per manuf. | Millipore | |
| α-HGF | nAb, quant-IB | Goat | HGF | Per manuf. | R&D Systems | |
| α-IFN-γ | nAb | Mouse | IFN-γ | Per manuf. | Millipore | |
| α-IGF-1 | nAb, quant-IB | Mouse | IGF-1 | Per manuf. | Millipore | |
| α-IL-6 | nAb | Mouse | IL-6 | Per manuf. | Millipore | |
| α-MCP-1 | nAb | Rabbit | MCP-1 | Per manuf. | Millipore | |
| α-TGF-β | nAb | Rabbit | TGF-βs (see Notes) | Per manuf. | R&D Systems | Detects β1-3 |
| α-TP53 | IB-Ab | Mouse | TP53 | 250 ng/mL | Cell Signaling (Beverly, MA) | |
| α-RasGAP | IB-Ab | Rabbit | RasGAP | 1:5000 Serum Ab | Kazlauskas Lab | |
| α-VEGFR2 | IB-Ab | Mouse | VEGFR2 | 100 ng/mL | Cell Signaling | |
| α-pVEGFR2 | IB-Ab | Rabbit | Phospho-VEGFR2 | 200 ng/mL | Cell Signaling | |
| HRP-α-IgG | Secondary Ab | Goat | Primary IB-Abs | Per manuf. | Santa Cruz | |
| Isotype IgGs | Control Ab | Human | NA | 52 μg/μL (1.2 mg) | Santa Cruz | ‡ |
| VEGF | GF | Human | VEGFR 1/2, PDGFRs | 2–50 ng/mL | NCI Repository | |
| PDGF-A | GF | Human | PDGFRα | 2–50 ng/mL | PeproTech (Rocky Hills, NJ) | |
| PDGF-AB | GF | Human | PDGFRs α and αβ | 2–50 ng/mL | PeproTech | |
| PDGF-B | GF | Human | PDGFRs | 2–50 ng/mL | PeproTech | |
| PDGF-C | GF, quant-IB | Human | PDGFRs α and αβ | 2–50 ng/mL | R&D Systems | |
| PDGF-D | GF, quant-IB | Human | PDGFRβ, (-αβ) | 2–50 ng/mL | R&D Systems | |
| CTGF | GF, quant-IB | Human | M6P, TrkA, others? | 2–50 ng/mL | PeproTech | |
| EGF | GF | Human | EGFR, HER2/3 | 5–50 ng/mL | PeproTech | |
| FGF-2 | GF | Human | FGFR-1 | 2–50 ng/mL | PeproTech | |
| G-CSF | GF | Human | G-CSF-R | 5–75 ng/mL | PeproTech | |
| HGF | GF, quant-IB | Human | c-Met | 2–50 ng/mL | PeproTech | |
| IFN-γ | GF | Human | IFN-γR | 0.5–25 ng/mL | PeproTech | |
| IGF-1 | GF, quant-IB | Human | IGFR-1, IR | 2–50 ng/mL | PeproTech | |
| IL-6 | GF | Human | IL-6R | 0.2–10 ng/mL | NCI Repository | |
| MCP-1 | GF | Human | CCR2 | 10–100 ng/mL | PeproTech | |
| TGF-α | GF | Human | EGFR | 5–50 ng/mL | PeproTech | |
| TGF-β1 | GF | Human | TGF-βR | 2–20 ng/mL | PeproTech | |
| TGF-β2 | GF | Human | TGF-βR | 2–20 ng/mL | PeproTech | |
| TGF-β3 | GF | Human | TGF-βR | 2–20 ng/mL | PeproTech |
All growth factor and cytokine antibodies used in this study detect both human and rabbit orthologs.
GF, recombinant human growth factor/cytokine; HRP, horseradish peroxidase; IB-Ab, Western immunoblot analysis antibody; Per manuf., set according to manufacturer's instructions; nAb, neutralizing antibody; PlGF, placenta growth factor; quant-IB, antibody used in quantitative Western immunoblot analysis or ligand used as a purified standard in quantitative Western immunoblot analysis; sR-Decoy, soluble receptor decoy (also considered ligand traps).
Values pertain to in vitro dosing, with the exception of values in parentheses, which indicate in vivo dosing amounts (ie, for intravitreal injections in experimental models).
Aflibercept is a fusion protein consisting of the extracellular domains of VEGFRs 1 and 2.
Amount used is equimolar to experimental treatment (with aflibercept).