Anti-CD20 (Rituximab biosimilar - IgG1fut isotype)

Non-fucosylated human IgG1 monoclonal antibody (mAb) against human CD20

Anti-hCD20-hIgG1fut features the variable region of rituximab and the constant region of a non-fucosylated, human IgG1 isotype. Rituximab is a mouse/human chimeric monoclonal antibody that targets the CD20 antigen found on the surface of malignant and normal B lymphocytes. The binding of rituximab to CD20 results in cell destruction through different mechanisms including direct signaling of apoptosis, complement activation, and cell-mediated cytotoxicity.

Rituximab has been approved by the FDA to treat various lymphoid malignancies, including B-cell non-Hodgkin's lymphoma and B-cell chronic lymphocytic leukemia.

Anti-hCD20-hIgG1fut is a non-fucosylated antibody. The absence of the fucose residue from the N-glycans of IgG-Fc results in dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) without any detectable change in complement-dependent cytotoxicity (CDC) or antigen-binding capability [1, 2].

Anti-hCD20-hIgG1fut was generated by recombinant DNA technology. It has been produced in Chinese hamster ovary (CHO) cells that are deficient for fucosylation and purified by affinity chromatography with protein G.

More isotypes of this antibody are available and can be used for comparison of biological activities such as ADCC (see below or in the 'upon request' section). InvivoGen also provides type I and type II anti-hCD20 mAbs featuring the variable regions of Ofatumumab or Obinutuzumab, with either native or engineered Fc regions.

References:

1. Yamane-Ohnuki N. & Satoh M., 2009. Production of therapeutic antibodies with controlled fucosylation. corresponding MAbs. 1(3): 230–236.
2. Mizushima T., 2011. Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. Genes Cells. 16(11): 1071–1080.

Figures

Comparison of ADCC potency for native and engineered anti-human CD20 antibody isotypes: Raji-Null cells were incubated with gradient concentrations of Anti-hCD20 or Anti-β-galactosidase (β-gal) mAbs for 1 hour. Jurkat-Lucia™ NFAT-CD16 effector cells were then co-incubated with targets cells for 6 hours. NFAT activation, reflecting the induced ADCC response, was assessed by determining Lucia luciferase activity in the supernatant using QUANTI-Luc™. Percentages of the maximal response normalized to the IgG1 isotype are shown

Specifications

Target: Human CD20

Clonality: Monoclonal antibody

Clone: Rituximab (Anti-hCD20-hIgG1, kappa)

Isotype: Human IgG1fut, kappa

Control: Human IgG1fut

Source: CHO cells

Formulation: 0.2 µm filtered solution in 68 mM sodium phosphate buffer (pH 7.4) with 91 mM glycine, 5% w/v saccharose, and stabilizing agents

Purity: Purified by affinity chromatography with protein G

Tested application: Flow cytometry and ADCC

Quality control:

• Binding of Anti-hCD20-hIgG1fut to human CD20 has been tested using flow cytometry.
• The complete sequence of this antibody has been verified.
• The absence of bacterial contamination (e.g. lipoproteins and endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.

Contents

Anti-hCD20-hIgG1fut purified monoclonal antibody is provided azide-free and lyophilized. It is available in two quantities:

• hcd20-mab13: 100 µg
• hcd20-mab13-03: 3 x 100 µg

Product is shipped at room temperature.

 Upon receipt, store lyophilized antibody at -20°C.