Anti-hCD20-hIgG4 (S228P)

Raji-Null Cells Unit size Cat. code Docs Qty Price
Human lymphoblast cells - ADCC CD19/CD20/Control Target Cells
3-7 x 10e6 cells
Anti-hCD20-hIgG4 (S228P) Unit size Cat. code Docs Qty Price
Engineered monoclonal human IgG4 antibody against human CD20
100 µg

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Engineered monoclonal human IgG4 antibody against human CD20

Effector functions of mAb isotypes targeting CD20

Anti-hCD20-hIgG4 (S228P) features the variable region of rituximab and a mutated constant region of the human IgG4 isotype reported to reduce Fab-arm exchange [1].

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-hIgG4 (S228P) is an engineered human IgG4 isotype. IgG4 antibodies display low antibody-dependent cell-mediated cytotoxicity (ADCC) and no complement-dependent cytotoxicity (CDC).

IgG4s are dynamic molecules that exchange Fab arms by swapping a heavy chain and attached light chain (half molecule) with a heavy-light chain pair from another molecule, resulting in bispecific antibodies [1, 2].

IgG4 molecules thereby lose their ability to cross-link antigen and to form immune complexes under most conditions [2]. Thus, mutations that prevent Fab-arm exchange in vivo should be considered when designing therapeutic IgG4.

The constant region of Anti-hCD20-hIgG4 (S228P) contains the S228P mutation which reduces Fab-arm exchange by stabilizing the disulfides in the core-hinge of the IgG4 molecules [1].

Anti-hCD20-hIgG4 (S228P) was generated by recombinant DNA technology. It has been produced in CHO cells and purified by affinity chromatography with protein G.



1. Labrijn AF. et al., 2009. Therapeutic IgG4 antibodies engage in Fab-arm exchange with endogenous human IgG4 in vivo. Nat Biotechnol. 27(8):767-71.
2. van der Neut Kolfschoten M. et al., 2007. Anti-inflammatory activity of human IgG4 antibodies by dynamic Fab arm exchange. Science. 317(5844):1554-7.


ADCC assay using various anti-human CD20 (Rituximab) antibody isotypes and Raji-Null target cells
ADCC assay using various anti-human CD20 (Rituximab) antibody isotypes and Raji-Null target cells

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

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Clonality: Engineered monoclonal antibody

Specificity: Targets cells expressing human CD20

Isotype: Human IgG4, kappa

Application :
Anti-hCD20-hIgG4 (S228P) can be compared with anti-hCD20-hIgG4 to study the impact of Fab-arm exchange.

Quality control

  • The absence of bacterial contamination (e.g. endotoxins and peptidoglycans) is controlled using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells.
  • The recognition of human CD20 with this antibody is tested using flow cytometry.
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  • 100 µg purified anti-hCD20-hIgG4 (S228P) antibody, provided azide-free and lyophilized.

room temperature Product is shipped at room temperature.

store Store lyophilized antibody at -20°C.

stability Lyophilized product is stable for 1 year.

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