Anti-CD20 (Obinutuzumab biosimilar - IgG1fut isotype)

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

Anti-hCD20-Ga-hIgG1fut is the biosimilar of the clinical antibody obinutuzumab. Anti-hCD20-Ga-hIgG1fut features the constant region (Fc) of a non-fucosylated (fut), human IgG1 isotype and the variable region of obinutuzumab. Obinutuzumab (formerly named afutuzumab or GA101) targets the human CD20 antigen found on the surface of normal and malignant B lymphocytes.

Obinutuzumab is a type II (second generation) non-fucosylated, fully humanized IgG1 monoclonal antibody with improved therapeutic efficacy compared with previously developed type I (first generation) anti-hCD20 mAbs, such as Rituximab and Ofatumumab. Obinutuzumab has been approved by the FDA to treat chronic lymphocytic leukemia [1-3].

The absence of fucose residues from the Fc N-glycans results in dramatic enhancement of antibody-dependent cellular cytotoxicity (ADCC) without any detectable change in complement-dependent cytotoxicity (CDC) or antigen-binding capability [4].

InvivoGen’s Anti-hCD20-Ga-hIgG1fut is generated by recombinant DNA technology, produced in Chinese hamster ovary (CHO) cells deficient for fucosylation, and purified by affinity chromatography (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 anti-hCD20 mAbs featuring the variable regions of Rituximab or Ofatumumab, with either native or engineered Fc region.

 Read our review on Antibody Isotypes

References:

1. Gagez AL. & Cartron G., 2014. Obinutuzumab: a new class of anti-CD20 monoclonal antibody. Curr Opin Oncol. 26(5):484-91.
2. Smolej L., 2014. Targeted treatment for chronic lymphocytic leukemia: clinical potential of obinutuzumab. Pharmgenomics Pers Med. 8:1-7.
3. Winiarska M. et al., 2011. Molecular mechanisms of the antitumor effects of anti-CD20 antibodies. Front Biosci (Landmark Ed). 16:277-306. 4. Mizushima T., 2011. Structural basis for improved efficacy of therapeutic antibodies on defucosylation of their Fc glycans. Genes Cells. 16(11): 1071–1080.

Figures

Binding of Anti-hCD20-Ga-hIgG1fut mAb to hCD20. EL4-hCD20 cells were incubated for 30 minutes at 4°C with 125 ng of Anti-hCD20-Ga-hIgG1fut or an isotype control. Subsequently, a secondary PE-labeled antibody was added and incubated at 4°C for 30 minutes. The binding affinity was assessed using flow cytometry.

Specifications

Target: Human CD20

Clonality: Monoclonal antibody

Clone: Obinutuzumab (Anti-hCD20-Ga-hIgG1fut, kappa)

Isotype: Human IgG1fut, kappa

Control: Human IgG1fut

Source: CHO cells (fucosylation-deficient)

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

Quality control:

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

• hcd20ga-mab13: 100 µg
• hcd20ga-mab13-03: 3 x 100 µg

Anti-hCD20-Ga-hIgG1fut is shipped at room temperature.

Store lyophilized antibody at -20 °C.

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