Detection of Antidrug Antibodies Against Human Therapeutic Antibodies Lacking Fc-Effector Functions by Usage of Soluble Fcγ Receptor I

Wessels, Uwe; Poehler, Alexander; Moheysen-Zadeh, Miriam; Zadak, Markus; Staack, Roland F.; Umaña, Pablo; Heinrich, Joachim; Stubenrauch, Kay

doi:10.4155/bio-2016-0182

Cited by 8 publications

(15 citation statements)

References 21 publications

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“…As expected [12], hsFcγRI binding to IgG1 bearing PG, LALA or PGLALA mutations was almost completely eliminated, whereas a strong signal was observed for human wtIgG1 (Figure 2, lower panel). Binding to wtIgG4 was weak in agreement to already published data [21,22].…”

Section: Specificity Of Anti-pg Mab and Hsfcγrimentioning

confidence: 57%

“…The improved assay format presented in this report can complement the bridging assay in immunogenicity assessment of therapeutic mAbs bearing the PG-substitution in the Fc region, which are used in several clinical development projects [12,14,15]. The assay principle, which, in similar design has been previously described for the preclinical assessment of drug-ADA complexes [11,[16][17][18], involves the capture of a drug-ADA complex by an immobilized antibody that is specific for the PG-mutagenized Fc region.…”

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confidence: 99%

“…In this assay, the secondary signal detection provides evidence that the assay signal is caused by an IgG1 antibody -a clear advantage over the bridging assay, which would detect any entity that is able to bind two drug molecules, such as an oligomeric soluble target. The hsFcγRI-based assay reliably detected human and Cynomolgus monkey IgG-ADAs against therapeutic monoclonal antibodies (mAbs) with suppressed Fc effector functions, such as those bearing Pro329Gly (PG) or Leu234Ala and Leu235Ala (LALA) substitutions in the Fc region [12][13][14]. The assay requires only one free ADA Fab fragment, as opposed to two in the bridging format, and hence it was less susceptible to the presence of residual drug.…”

mentioning

confidence: 99%

“…Bound immune complexes are detected using hsFcγRI ( Figure 1C). Since the hsFcγRI does not bind to the PG-modified Fc region of the drug ( Figure 1D; [12]) and the anti-PG capture antibody, in turn cannot bind to the wild-type IgG (wtIgG; Figure 1E), the detection is specific for the drug-ADA complex. We therefore expect that the assay is intrinsically insensitive to even high drug concentrations in the matrix.…”

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confidence: 99%

“…We recently described a novel assay with an orthogonal detection method based on digoxigenin (Dig) labeled ectodomain (extracellular ligand-binding domain) of the human Fcγ receptor I (hsFcγRI; see Figure 1B) [12]. In this assay, the secondary signal detection provides evidence that the assay signal is caused by an IgG1 antibody -a clear advantage over the bridging assay, which would detect any entity that is able to bind two drug molecules, such as an oligomeric soluble target.…”

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confidence: 99%

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Novel Drug and Soluble Target Tolerant Antidrug Antibody Assay for Therapeutic Antibodies Bearing The P329G Mutation

et al. 2017

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Aim:Bridging immunoassays for detection of antidrug antibodies (ADAs) are typically susceptible to high concentrations of residual drug. Sensitive drug-tolerant assays are, therefore, needed. Materials & methods: An immune complex assay to detect ADAs against therapeutic antibodies bearing Pro329Gly mutation was established. The assay uses antibodies specific for the Pro329Gly mutation for capture and human soluble Fcγ receptor for detection. Results: When compared with a bridging assay, the new assay showed similar precision, high sensitivity to IgG1 ADA and dramatically improved drug tolerance. However, it was not able to detect early (IgM-based) immune responses. Conclusion: Applied in combination with a bridging assay, the novel assay serves as orthogonal assay for immunogenicity assessment and allows further characterization of ADA responses.

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Section: Specificity Of Anti-pg Mab and Hsfcγrimentioning

confidence: 57%

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confidence: 99%

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confidence: 99%

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confidence: 99%

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confidence: 99%

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Novel Drug and Soluble Target Tolerant Antidrug Antibody Assay for Therapeutic Antibodies Bearing The P329G Mutation

et al. 2017

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Fc-independent neutralization of SARS-CoV-2 by recombinant human monoclonal antibodies

Noy-Porat

Edri

Alcalay

et al. 2021

Preprint

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The use of passively-administered neutralizing antibodies is a promising approach for the prevention and treatment of SARS-CoV-2 infection. Antibody-mediated protection may involve immune system recruitment through Fc-dependent activation of effector cells and the complement system. However, the role of Fc-mediated functions in the efficacious in vivo neutralization of SARS-CoV-2 is not yet clear. Delineating the role this process plays in antibody-mediated protection will have a great impact on the design of such therapeutics. Here, the Fc of two highly potent SARS-CoV-2 neutralizing human monoclonal antibodies, targeting distinct domains of the spike, was engineered to abrogate their Fc-dependent functions. The protective activity of these antibodies was tested against lethal SARS-CoV-2 infections in K18-hACE2 transgenic mice, both before or two days post-exposure in comparison to their original, Fc-active antibodies. Antibody treatment with both Fc-variants similarly rescued the mice from death, reduced viral load and prevented signs of morbidity. In addition, surviving animals developed a significant endogenous immune response towards the virus. Taken together, this work provides important insight regarding the contribution of Fc-effector functions in antibody-mediated protection, which should aid in future design of effective antibody-based therapies.

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Conceptual Approaches to Modulating Antibody Effector Functions and Circulation Half-Life

2019

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Antibodies and Fc-fusion antibody-like proteins have become successful biologics developed for cancer treatment, passive immunity against infection, addiction, and autoimmune diseases. In general these biopharmaceuticals can be used for blocking protein:protein interactions, crosslinking host receptors to induce signaling, recruiting effector cells to targets, and fixing complement. With the vast capability of antibodies to affect infectious and genetic diseases much effort has been placed on improving and tailoring antibodies for specific functions. While antibody:antigen engagement is critical for an efficacious antibody biologic, equally as important are the hinge and constant domains of the heavy chain. It is the hinge and constant domains of the antibody that engage host receptors or complement protein to mediate a myriad of effector functions and regulate antibody circulation. Molecular and structural studies have provided insight into how the hinge and constant domains from antibodies across different species, isotypes, subclasses, and alleles are recognized by host cell receptors and complement protein C1q. The molecular details of these interactions have led to manipulation of the sequences and glycosylation of hinge and constant domains to enhance or reduce antibody effector functions and circulating half-life. This review will describe the concepts being applied to optimize the hinge and crystallizable fragment of antibodies, and it will detail how these interactions can be tuned up or down to mediate a biological function that confers a desired disease outcome.

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Detection of Antidrug Antibodies Against Human Therapeutic Antibodies Lacking Fc-Effector Functions by Usage of Soluble Fcγ Receptor I

Abstract: The hsFcγRI-based ADA assay can serve as alternative screening assay or as orthogonal confirmation method for preclinical and clinical immunogenicity testing of IgG therapeutics lacking Fc effector functions.

Cited by 8 publications

References 21 publications

Novel Drug and Soluble Target Tolerant Antidrug Antibody Assay for Therapeutic Antibodies Bearing The P329G Mutation

Novel Drug and Soluble Target Tolerant Antidrug Antibody Assay for Therapeutic Antibodies Bearing The P329G Mutation

Fc-independent neutralization of SARS-CoV-2 by recombinant human monoclonal antibodies

Conceptual Approaches to Modulating Antibody Effector Functions and Circulation Half-Life

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