Anti-human IL20 Recombinant Antibody (2A11) (TAB-562LC) (CAT#: TAB-562LC)

It is a recombinant antibody targeting the human interleukin 20, derived from human. It specifically binds to human IL-20 and can be potentially used in the research of autoimmune and autoinflammatory disorders. This antibody has been detected using Enzyme-linked immunosorbent assay, Surface plasmon resonance and Neutralization assay.


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ELISA

Figure 1 Affinity of Patient-Derived mAbs

Figure 1 Affinity of Patient-Derived mAbs

Binding determined by LIPS of APS1/APECED-derived mAbs and of germline counterparts to IFNa2, IFNa8, IFNa14 (19D11, 50E11, and 26B9), IL22 (35G11 and 30G1), and IL20 (20A10 and 2A11). Binding to immobilized IL17F (17E3 and 9A2) was determined by ELISA.

Meyer, S., Woodward, M., Hertel, C., Vlaicu, P., Haque, Y., Kärner, J.,... & Metsküla, K. (2016). AIRE-deficient patients harbor unique high-affinity disease-ameliorating autoantibodies. Cell, 166(3), 582-595.

ELISA

Figure 2 ELISA determination of binding of exemplary anti-IL-20 antibodies to IL-20 constructs fused either N-terminally (gl IL-20) or C-terminally (g2 IL-20) to Gaussia luciferase.

Figure 2 ELISA determination of binding of exemplary anti-IL-20 antibodies to IL-20 constructs fused either N-terminally (gl IL-20) or C-terminally (g2 IL-20) to Gaussia luciferase.

A: 2A11; B: 6E11; C: 6H2; D: 7D1; E: 20A10; F: A non-IL20 specific monoclonal antibody is used as control for non-specific binding. Antibodies 2A11, 6E11 and 20A10 bind with a high affinity to both IL-20 constructs, in contrast remaining antibodies 6H2 and 7D1 bind with a lower affinity to g2 IL-20.

Neut

Figure 3 Human-derived anti-IL-20 monoclonal antibodies neutralize recombinant IL-20-mediated STAT3 activation in HEK 293T MSR cells transiently expressing Type I and Type II IL-20 receptors.

Figure 3 Human-derived anti-IL-20 monoclonal antibodies neutralize recombinant IL-20-mediated STAT3 activation in HEK 293T MSR cells transiently expressing Type I and Type II IL-20 receptors.

Either cells were left untreated or stimulated with recombinant human or murine IL-20 in the absence of antibodies or in the presence of human-derived IL-20 monoclonal antibodies or a human control IgG as indicated (hulgG). Cell lysates were subjected to SDS-PAGE and pSTAT3 levels were visualized in Western blots. Total STAT3, alpha-tubulin and IL-20RB levels serve as loading control. Antibody concentration: 5 μg/ml. A: Control: rhIL-20 and rmlL-20 induce the dose-dependent phosphorylation of STAT3 in HEK 293T MSR cells transiently expressing Type I or Type II IL-20 receptors. Detection of total and phosphorylated STAT3 and IL-20RB-Myc-DDK levels. B: Stimulation with rhIL-20 (10 ng/ml) or rmIL-20 (25 ng/ml). Exemplary antibodies 2A11, 20A10 and 7D1 neutralize rhIL-20, whereas only 20A10 also potently neutralizes rmIL-20.

Neut

Figure 4 Human-derived anti-IL-20 monoclonal antibodies neutralize rhIL-20-mediated induction of KZ136-NLuc reporter constructs in HEK 293T MSR cells transiently transfected with IL-20 receptors and KZ136-NLuc.

Figure 4 Human-derived anti-IL-20 monoclonal antibodies neutralize rhIL-20-mediated induction of KZ136-NLuc reporter constructs in HEK 293T MSR cells transiently transfected with IL-20 receptors and KZ136-NLuc.

Cells were stimulated with 120 ng/ml rhIL-20 or 200 ng/ml rmIL-20 in the presence of human-derived IL-20 monoclonal antibodies or of a control human IgG (hulgG) as indicated. rhlL-20 IC50analysis of exemplary antibodies 20A10, 7D1 and 2A11.

Neut

Figure 5 IC50 analysis of human-derived anti-IL-20 monoclonal antibodies in the chemiluminescent cellular binding assay.

Figure 5 IC50 analysis of human-derived anti-IL-20 monoclonal antibodies in the chemiluminescent cellular binding assay.

HEK 293T MSR cells transiently expressing Type I or Type II IL-20 receptors were incubated with gl IL-20 supernatants in the presence of human-derived IL-20 monoclonal antibodies, A: 20A10, B: 7D1 and C: 2A11. D: IC50 values of exemplary antibodies 20A10, 7D1 and 2Al l.

ELISA

Figure 6 Cross-competition

Figure 6 Cross-competition

Experimental setup: 96 well microplates (Costar, USA) were coated with Rabbit-anti-GLuc specific antibody diluted 1/250 in PBS O/N at 4 °C. Plates were washed with PBS-T and blocked lh at room temperature with PBS containing 2 % BSA (Sigma, Buchs, Switzerland). 30 μL GLuc-IL-20 was added at a final concentration of 2x10⁶ LU/well and incubated for 2h at room temperature. The chimeric antibodies are premixed with the human competitor antibodies at a final concentration of 0.5 μg/ml vs. 3 μg/ml in 30 μL PBS and added to the plates. Upon incubation for 2 h at room temperature the plates were washed with PBS-T and the binding of the human-mouse chimeric antibodies was determined using a horseradish peroxidase conjugated goat anti-mouse IgG Fc-gamma specific antibody (Jackson Immuno Research, 1 :500 in 0.5 % BSA-PBS) followed by measurement of the HRP activity using a TMB substrate solution (Sigma, Buchs, Switzerland). The differential binding of exemplary human-derived anti-IL-20 monoclonal antibodies with human-mouse (hm) chimeric constructs to distinct binding sites was investigated in cross-competition experiments with 2A11 hm.

ELISA

Figure 7 Cross-reactivity-Determination and comparison of binding of exemplary human-derived anti-IL-20 monoclonal antibodies 6E11, 2A11 and 20A10 to mouse and human IL-20 by GLuc-sandwich ELISA.

Figure 7 Cross-reactivity-Determination and comparison of binding of exemplary human-derived anti-IL-20 monoclonal antibodies 6E11, 2A11 and 20A10 to mouse and human IL-20 by GLuc-sandwich ELISA.

SPR

Figure 8 SPR analysis

Figure 8 SPR analysis

Detailed analysis of the sensograms concerning the binding of human and mouse IL-20 to exemplary antibodies. A 1 : 1 binding kinetic was observed. The antigens were injected in concentrations of 1.5 nM, 3.125 nM, 6.25 nM, 12.5 nM, 25 nM, 50 nM and 100 nM. Calculated Affinities (KD values [M]) are indicated in the diagrams.

Figure 9 Mapping of the epitopes of the human anti-IL-20 antibodies.

Figure 9 Mapping of the epitopes of the human anti-IL-20 antibodies.

Binding of antibodies to full length antigens coupled to the microarray. The Y-axis indicated the fluorescence intensity (RFU) upon detection with a Cy5-conjugated secondary antibody.


Specifications

  • Host Species
  • Human
  • Type
  • Human antibody
  • Specificity
  • Human
  • Clone
  • 2A11
  • Applications
  • ELISA, Neut, SPR
  • Related Disease
  • Autoimmune and autoinflammatory disorder

Applications

  • Application Notes
  • This antibody has been tested for use in Enzyme-Linked Immunosorbent Assay, Neutralization and Surface Plasmon Resonance.

Target

  • Alternative Names
  • IL20; interleukin 20; IL-20; IL10D; ZCYTO10; interleukin-20; cytokine Zcyto10; four alpha helix cytokine

Product Notes

This is a product of Creative Biolabs' Hi-Affi™ recombinant antibody portfolio, which has several benefits including:

• Increased sensitivity
• Confirmed specificity
• High repeatability
• Excellent batch-to-batch consistency
• Sustainable supply
• Animal-free production

See more details about Hi-Affi™ recombinant antibody benefits.

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Human Antibody

CAT Product Name Application Type
TAB-H34 Anti-Human IL20 Recombinant Antibody (Fletikumab) IF, IP, Neut, FuncS, ELISA, FC, ICC IgG4 - kappa
TAB-561LC-F(E) Anti-human IL20 Fab Fragment (20A10) ELISA Human antibody
TAB-562LC-F(E) Anti-human IL20 Fab Fragment (2A11) ELISA Human antibody
TAB-563LC-F(E) Anti-human IL20 Fab Fragment (7D1) ELISA Human antibody
TAB-564LC-F(E) Anti-human IL20 Fab Fragment (6E11) ELISA Human antibody

Humanized Antibody

Neutralizing Antibody

CAT Product Name Application Type
NEUT-1324CQ Rat Anti-Il20 Recombinant Antibody (clone CBL626) Neut Rat IgG2b

Rabbit Monoclonal Antibody

CAT Product Name Application Type
MOR-4256 Rabbit Anti-IL20 Recombinant Antibody (clone SI336DS) ELISA Rabbit IgG

ADCC Enhanced Antibody

CAT Product Name Application Type
AFC-TAB-H34 Afuco™ Anti-IL20 ADCC Recombinant Antibody (Fletikumab), ADCC Enhanced IF, IP, Neut, FuncS, ELISA, FC ADCC enhanced antibody

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For Research Use Only. Not For Clinical Use.

For research use only. Not intended for any clinical use. No products from Creative Biolabs may be resold, modified for resale or used to manufacture commercial products without prior written approval from Creative Biolabs.

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