Afuco™ Anti-Human HLA-DR10 ADCC Recombinant Antibody (LYM-1), ADCC Enhanced (CAT#: AFC-347CL)

Anti-HLA-DR10 ADCC Enhanced Antibody (LYM-1) is an ADCC enhanced antibody produced by our Afuco™ platform. The monoclonal antibodies Lym-1 reactive with the cell surface of B-lymphocytes and derived tumors, have indicated significant anti-tumour activity against lymphoma cell lines.


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Figure 1 Confocal micrographs of washed live cells after incubation with chLym-1, tridentate (DvPLLCtPCbLB) or dimeric, tridentate ((DvPLLCtPCbPPP)2LLB) SHAL (left to right) in Raji (expressing; top row), and Jurkat's (nonexpressing; bottom row) cells.

Figure 1 Confocal micrographs of washed live cells after incubation with chLym-1, tridentate (DvPLLCtPCbLB) or dimeric, tridentate ((DvPLLCtPCbPPP)2LLB) SHAL (left to right) in Raji (expressing; top row), and Jurkat's (nonexpressing; bottom row) cells.

Lym-1 (left) showed Raji cell-surface membrane binding, whereas the tridentate (middle) and the dimeric, tridentate SHALs (right) showed intracellular Raji binding. There was no binding in Jurkat's cells. DAPI (blue) was used as the nuclear stain, and Alexa Fluor® (Invitrogen, Carlsbad, CA) (red) was used for the location of SHAL or Lym-1 in these merged laser images.

DeNardo, G. L., Natarajan, A., Hok, S., Mirick, G., DeNardo, S. J., Corzett, M., ... & Balhorn, R. (2008). Nanomolecular HLA-DR10 antibody mimics: a potent system for molecular targeted therapy and imaging. Cancer biotherapy & radiopharmaceuticals, 23(6), 783-795.

Figure 2 Binding of SHAL2 and Lym-1 antibody to B-cell lymphoma tissue from patients.

Figure 2 Binding of SHAL2 and Lym-1 antibody to B-cell lymphoma tissue from patients.

SHAL2 was preincubated with horseradish peroxidase-tagged streptavidin and binding to the cells in the tissue was detected using 3,3 ¶ -diaminobenzidine reagent. Lym-1 binding was detected with a biotinylated antimouse monoclonal antibody, followed by horseradish peroxidase-tagged streptavidin and 3,3-diaminobenzidine. A, SHAL 2 on large cell lymphoma. B, SHAL 2 on small cell lymphoma. C, Lym-1antibody on large cell lymphoma. D, Lym-1antibody on small cell lymphoma.

Balhorn, R., Hok, S., Burke, P. A., Lightstone, F. C., Cosman, M., Zemla, A., ... & DeNardo, S. J. (2007). Selective high-affinity ligand antibody mimics for cancer diagnosis and therapy: initial application to lymphoma/leukemia. Clinical Cancer Research, 13(18), 5621s-5628s.

Figure 3 Depletion of CD19-positive splenocytes by anti-CD20 and anti-HLA-DR antibodies.

Figure 3 Depletion of CD19-positive splenocytes by anti-CD20 and anti-HLA-DR antibodies.

Antibodies used were: Hu1D10, anti-CD20 antibody B1, anti-HLA-DR variant antibody Lym-1 and negative control HuFd79 (humanized antibody against herpes simplex virus). Similar results were obtained with an additional sample of 1D10 (+) splenocytes.

Kostelny, S. A., Link, B. K., Tso, J. Y., Vasquez, M., Jorgensen, B. H., Wang, H., ... & Weiner, G. J. (2001). Humanization and characterization of the anti-HLA-DR antibody 1D10. International journal of cancer, 93(4), 556-565.

Figure 4 Apoptosis of B-cells induced by anti-HLDR antibodies.

Figure 4 Apoptosis of B-cells induced by anti-HLDR antibodies.

Raji (a), Priess (b), Daudi (c) and splenic B-cell (d) death induced by various anti-HLA-DR antibodies and an anti-CD20 antibody. Antibodies used were 1D10 (murine IgG2a/kappa), Hu1D10 (human IgG1/kappa), F(ab′)2 form of Hu1D10 (Hu1D10-F(ab′)2), pan-anti-HLA-DR antibodies L234 and L227 (both are murine IgG2a/kappa), anti-CD20 antibody B1 (murine IgG2a/kappa), anti-HLA-DR variant antibody Lym-1 (murine IgG2a/kappa), and isotype control antibodies UPC10 (murine IgG2a/kappa) and HuFd79 (humanized antibody against herpes simplex virus, human IgG1/kappa).

Kostelny, S. A., Link, B. K., Tso, J. Y., Vasquez, M., Jorgensen, B. H., Wang, H., ... & Weiner, G. J. (2001). Humanization and characterization of the anti-HLA-DR antibody 1D10. International journal of cancer, 93(4), 556-565.

Figure 5 Two-color fluorescence-activated cell sorter (FACS) analysis of blood lymphocytes from a Hu1D10 positive donor.

Figure 5 Two-color fluorescence-activated cell sorter (FACS) analysis of blood lymphocytes from a Hu1D10 positive donor.

Blood samples were stained separately with fluorescein isothiocyanate (FITC)-conjugated Hu1D10, FITC-conjugated Lym-1 or FITC-conjugated L243 (anti-HLA-DR). Phycoerythrin (PE)-conjugated anti-CD19 antibody was added to all samples. White blood cells in each sample were processed, and the lymphocytes were analyzed by FACScan for 2-color immunofluorescence. Lymphocytes in the left panel were stained with anti-CD19 and Hu1D10, in middle panel with anti-CD19 and Lym-1, and in the right panel with anti-CD19 and L243. The number in each quadrant represents the percentage of cells in that area.

Kostelny, S. A., Link, B. K., Tso, J. Y., Vasquez, M., Jorgensen, B. H., Wang, H., ... & Weiner, G. J. (2001). Humanization and characterization of the anti-HLA-DR antibody 1D10. International journal of cancer, 93(4), 556-565.

Figure 6 Western blot analysis was used to compare the precolumn, flow-through, and column-purified Lym-1 antigen samples of Raji cell lysate.

Figure 6 Western blot analysis was used to compare the precolumn, flow-through, and column-purified Lym-1 antigen samples of Raji cell lysate.

Not-heated/not-reduced samples of lane 1 pre-cleared Raji cell lysate, lane 2 Lym-1 column flow-through, and lane 3 immunoaffinity-purified Lym-1 antigen were separated on SDSPAGE gels, Western blotted and stained with A Lym-1, B anti-DRβ, C anti-DR52, and D anti-DR10. Lym-1 and anti-DR10 both react with the Raji lysate and the immunoaffinity-purified Lym-1 antigen, but not with the affinity column flow-through. Anti-DRβ and anti-DR52 mAb react with all three samples, though anti-DR52 reacts only weakly with the affinity-purified Lym-1 antigen.

Rose, L. M., Gunasekera, A. H., DeNardo, S. J., DeNardo, G. L., & Meares, C. F. (1996). Lymphoma-selective antibody Lym-1 recognizes a discontinuous epitope on the light chain of HLA-DR10. Cancer Immunology, Immunotherapy, 43(1), 26-30.


Specifications

  • Host Species
  • Mouse
  • Derivation
  • Mouse
  • Type
  • ADCC enhanced antibody
  • Species Reactivity
  • Human
  • Related Disease
  • Non-Hodgkin's Lymphoma

Product Property

  • Purity
  • >95% as judged by SDS-polyacrylamide gel electrophoresis
  • Storage
  • 4°C or -20°C, avoid repeated freezing and thawing.

Target

  • Alternative Names
  • HLA-DR10; Human Leukocyte Antigen-DR10; DR10

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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