Tandemly repeated Fc domain augments binding avidities of antibodies for Fcγ receptors, resulting in enhanced antibody-dependent cellular cytotoxicity

Nagashima, Hiroaki; Tezuka, T.; Tsuchida, W.S.; Maeda, Hiroaki; Kohroki, Junya; Masuho, Yasuhiko

doi:10.1016/j.molimm.2008.02.003

Cited by 23 publications

(17 citation statements)

References 35 publications

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“…In addition, Nagashima et al . 35 developed an antibody with three tandemly repeated Fc domains at the c-terminus, and found that it exhibited about 100 times greater avidity than wild type antibody for Fc receptors (FcRIIA, FcRIIB and FcRIIIA). Therefore, the amount of antibody that the protein G C2 domain can trap appears to be dramatically increased after it is rendered into a repetitive polymer.…”

Section: Discussionmentioning

confidence: 99%

Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays

Hao

Chen

et al. 2017

Sci Rep

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The sensitivities of solid-phase immunoassays are limited by the quantity of detection antibodies bound to their antigens on the solid phase. Here, we developed a poly-protein G-expressing bacterium as an antibody-trapping microparticle to enhance the signals of immunoassays by increasing the accumulation of detection antibodies on the given antigen. Eight tandemly repeated fragment crystallisable (Fc) binding domains of protein G were stably expressed on the surface of Escherichia coli BL21 cells (termed BL21/8G). BL21/8G cells showed a higher avidity for trapping antibodies on their surface than monomeric protein G-expressing BL21 (BL21/1G) cells did. In the sandwich enzyme-linked immunosorbent assay (ELISA), simply mixing the detection antibody with BL21/8G provided a detection limit of 6 pg/mL for human interferon-α (IFN-α) and a limit of 30 pg/mL for polyethylene glycol (PEG)-conjugated IFN-α (Pegasys), which are better than that of the traditional ELISA (30 pg/mL for IFN-α and 100 pg/mL for Pegasys). Moreover, the sensitivity of the Western blot for low-abundance Pegasys (0.4 ng/well) was increased by 25 folds upon mixing of an anti-PEG antibody with BL21/8G cells. By simply being mixed with a detection antibody, the poly-protein G-expressing bacteria can provide a new method to sensitively detect low-abundance target molecules in solid-phase immunoassays.

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Section: Discussionmentioning

confidence: 99%

Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays

Hao

Chen

et al. 2017

Sci Rep

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“…This is supported by a recent study with linearly connected Fc domains, which showed by ELISA assay augmented binding affinities to FcγRIIA, FcγRIIB and FcγRIIIA in the absence of the antigen. 46 The change in the FcγRI-binding was to much lesser extent. Although the sensorgrams shown in Figure 10 could fit the "conformational change" model provided in Biacore software reasonably well, we did not report the fitting results since a binding constant would be ill defined in this case due to the heterogeneity in size of the complexes.…”

Section: Discussionmentioning

confidence: 99%

Dimers and multimers of monoclonal IgG1 exhibit higher in vitro binding affinities to Fcγ receptors

Luo

Raso

et al. 2009

mAbs

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“…Other mAb frameworks utilizing Fc multimerization strategies have shown augmented effector functions including ADCP, ADCC, and opsonophagocytic killing . These tandem Fc proteins are similar to the 2Fc mAb in terms of size, biochemical properties, and improvement of FcγR binding.…”

Section: Discussionmentioning

confidence: 99%

Design and characterization of novel dual Fc antibody with enhanced avidity for Fc receptors

et al. 2019

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Monoclonal antibodies (mAbs) have become an important class of therapeutics, particularly in the realm of anticancer immunotherapy. While the two antigen‐binding fragments (Fabs) of an mAb allow for high‐avidity binding to molecular targets, the crystallizable fragment (Fc) engages immune effector elements. mAbs of the IgG class are used for the treatment of autoimmune diseases and can elicit antitumor immune functions not only by several mechanisms including direct antigen engagement via their Fab arms but also by Fab binding to tumors combined with Fc engagement of complement component C1q and Fcγ receptors. Additionally, IgG binding to the neonatal Fc receptor (FcRn) allows for endosomal recycling and prolonged serum half‐life. To augment the effector functions or half‐life of an IgG1 mAb, we constructed a novel “2Fc” mAb containing two Fc domains in addition to the normal two Fab domains. Structural and functional characterization of this 2Fc mAb demonstrated that it exists in a tetrahedral‐like geometry and retains binding capacity via the Fab domains. Furthermore, duplication of the Fc region significantly enhanced avidity for Fc receptors FcγRI, FcγRIIIa, and FcRn, which manifested as a decrease in complex dissociation rate that was more pronounced at higher densities of receptor. At intermediate receptor density, the dissociation rate for Fc receptors was decreased 6‐ to 130‐fold, resulting in apparent affinity increases of 7‐ to 42‐fold. Stoichiometric analysis confirmed that each 2Fc mAb may simultaneously bind two molecules of FcγRI or four molecules of FcRn, which is double the stoichiometry of a wild‐type mAb. In summary, duplication of the IgG Fc region allows for increased avidity to Fc receptors that could translate into clinically relevant enhancement of effector functions or pharmacokinetics.

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Tandemly repeated Fc domain augments binding avidities of antibodies for Fcγ receptors, resulting in enhanced antibody-dependent cellular cytotoxicity

Cited by 23 publications

References 35 publications

Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays

Poly-protein G-expressing bacteria enhance the sensitivity of immunoassays

Dimers and multimers of monoclonal IgG1 exhibit higher in vitro binding affinities to Fcγ receptors

Design and characterization of novel dual Fc antibody with enhanced avidity for Fc receptors

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