Anti-hCD20-hIgG1NQ

Non-glycosylated monoclonal human IgG1 antibody against human CD20

Anti-hCD20-hIgG1NQ features the variable region of rituximab and a mutated constant region of the 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. 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 for the treatment of various lymphoid malignancies, including B-cell non-Hodgkin's lymphoma and B-cell chronic lymphocytic leukemia.

Anti-hCD20-hIgG1NQ contains a N-glycosylation mutation of the constant region of the human IgG1 where potential asparagine (N) glycosylation sites are substituted by glutamine (Q) residues resulting in the production of a non-glycosylated antibody. Glycosylation of an antibody has no effect on antigen binding but is essential for Fc receptor-mediated activity [1].
In non-glycosylated antibodies the effector mechanisms mediated through the Fc receptors types (FcgRI, FcgRII, FcgRIII) and the C1q component of complement are severely compromised or ablated [2].
Anti-hCD20-hIgG1NQ was generated by recombinant DNA technology. It has been produced in CHO cells and purified by affinity chromatography with protein G.

Réferences:

1. Arnold J. et al., 2007. The impact of glycosylation on the biological function and structure of human immunoglobulins. Annu Rev Immunol 25:21-50.
2. Jefferis R., 2009. Glycosylation as a strategy to improve antibody-based therapeutics. Nat Rev Drug Discov. 8(3):226-34.

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

Clonality: Non-glycosylated Monoclonal antibody

Specificity: Targets cells expressing human CD20

Isotype: Human IgG1, kappa

Application :
Anti-hCD20-hIgG1NQ can be compared with anti-hCD20-hIgG1 to study the impact of effector functions.

Quality control

• The absence of bacterial contamination (e.g. endotoxins and peptidoglycans) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.
• The recognition of human CD20 with this antibody has been tested using flow cytometry.

Contents

• 100 µg purified anti-hCD20-hIgG1NQ antibody, provided azide-free and lyophilized.

Product is shipped at room temperature.

Store lyophilized antibody at -20°C.

Lyophilized product is stable for 1 year