Book a Meeting

Fc Engineering Service for Effector Function Stimulation

The antibody Fc domain engages with a small family of Fc receptors expressed on immune system cells and other cells to stimulate the rich diversity of positive and negative cell-mediated effector functions. The emergence of monoclonal antibodies for the treatment of various pathological conditions has provided more insights into the biology of Fc receptors and proposed novel strategies for the development of improved therapies using Fc receptor interactions. Although most therapeutic IgGs approved to date retain the natural IgG Fc domain, the knowledge gained in the past few decades provides opportunities for the Fc design of tailored and more effective immunotherapies that have shown fewer side effects and longer half-life.

Creative Biolabs keeps track of the latest advances made in the design of biologics that modulate or exploit Fc receptor-IgG interactions, and are committed to developing the optimal Fc mutation strategy to support the innovative drugs currently being studied in clinical trials, and fine-tune them to achieve premium effector functions (ADCC, ADCP, and CDC).

Fc functions in stimulating the immune system

IgG Fc receptors (such as FcgRI, FcgRIIa and FcgRIIIa) possess the classic cytoplasmic ITAM (immunoreceptor tyrosine-based activation motif) signal transduction domain, and can stimulate immune effector cell activation and subsequent antibody-dependent cell activation (e.g. mediated cytotoxicity (ADCC) or phagocytosis (ADCP). Except for FcgRI, most FcgRs have low affinity for ligands (> 1 mM). This is of vital importance for the development of Fc engineered antibody therapy. Therefore, the weak Fc binding of several key Fc receptors facilitates the engineering of novel monomeric antibodies to achieve the desired therapeutic effect.

Through the modification of the Fc region, the binding of Fc to FcγRIIa on macrophages and FcγRIIIa on NK cells can be enhanced, and these cells can be recruited more effectively, thereby killing tumor cells. Recently, it has been shown in FcgR humanized mice that the Fc binding of human FcgRIIa not only stimulates macrophage-mediated ADCP, but FcgRIIa is also a necessary condition and sufficient for inducing strong anti-tumor cell immunity mediated by dendritic cells and T cells. Such results raise the exciting prospect of coupling Fc-enhanced cytotoxic tumor-targeting antibodies with more non-selective immunostimulants such as checkpoint blockade.

Strategies to regulate the immune response mediated by FcgR. Fig.1 Strategies to regulate the immune response mediated by FcgR. (Sondermann, 2016)

By manipulating the recruitment of activated and inhibitory receptors, such as through engineered antibodies, a treatment plan can be tailored to the optimal treatment for a specific indication.

Highlight clinical trials of engineered Fc to stimulate the immune system

Studies have reported that single mutations (S239D or I332E), double mutations (S239D/I332E) and triple mutations (S239D/I332E/A330L) in the Fc region can increase the affinity of Fc for FcγRIIa and FcγRIIIa, and the affinity for FcγRIIIa has increased to 169. Times. After these modifications, CD19 and CD96 antibodies all showed stronger ADCC activity. In addition, a point mutation G236A can selectively recognize FcγRIIa but not FcγRIIb, so it can significantly increase ADCP activity. Mutation of G236A and S239D/I332E can increase ADCC and ADCP activity at the same time.

Fc Engineering Isotype Intended Function Result
S239D/I332E or S239D/I332E/A330L Glyco-IgG1 Higher affinity to FcgRIIIa Enhance ADCC
G236A Glyco-IgG1 Higher affinity to FcgRIIa
F243L/R292P/Y300L/V3051/P396L Glyco-IgG1 Higher affinity to FcgRI
S298A/E333A/K334A Glyco-IgG1 Higher affinity to FcgRIIIa
N297A Glyco-IgG1 Afucosylated
G236A/S239D/I332E Glyco-IgG1 Increased FcγRIIa binding, Increased FcγRIIIa binding Enhance ADCP
K326W/E333S Glyco-IgG1 Increased C1q binding Enhance CDC
S267E/H268F/S324T Glyco-IgG1 Increased C1q binding
IgG1/IgG3 cross subclass Glyco-IgG1 Increased C1q binding
E345R/E430G/S440Y Glyco-IgG1 Hexamerization

Fig.2 Fc modification for activating the immune system.

XmAb5574 is an anti-CD19 antibody. It has obtained a stronger NK cell-mediated ADCC/ADCP effect through the transformation of S239D/I332E. It is used for the treatment of CLL and is currently in Phase I clinical trials. Similar to XmAb2513 (anti-CD30), JNJ-56022473 (anti-CD123), PF-04605412 (integrin α5β1) and BI 836826 (anti-CD37).

For more detailed information, please feel free to contact us or directly sent us an inquiry.


  1. Sondermann, P., & Szymkowski, D. E. (2016). Harnessing Fc receptor biology in the design of therapeutic antibodies. Current opinion in immunology, 40, 78-87.


Creative Biolabs provides luciferase-based ADCC assay. This Jurkat cell based assay is pioneered by Creative Biolabs, and the methodology is very well accepted by the field. See attached ADCC Reporter Assay Protocol for further details. 

All products and services are for Research Use Only. Do Not use in humans.


ADCC Assay WT vs AfucoTM Mabs  Visit


Antibody Fc Engineering: Towards Better Therapeutics  Visit


Creative Biolabs has established a team of customer support scientists ready to discuss ADCC/CDC optimization strategies, antibody production, bioinformatics analysis and other molecular biology/biotechnology issues.

  • *
  • *
  • *