Charge-mediated influence of the antibody variable domain on FcRn-dependent pharmacokinetics

Schoch, Angela; Kettenberger, Hubert; Mundigl, Olaf; Winter, Gerhard; Engert, Julia; Heinrich, Julia; Emrich, Thomas

doi:10.1073/pnas.1408766112

Cited by 172 publications

(245 citation statements)

References 46 publications

(62 reference statements)

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“…Some, but not all, of these differences can be ascribed to physiochemical characteristics of the variable sequences. 46 In addition, the hinge domain that links the Fab to Fc domain also plays a role in the mFcRn-IgG affinity. 47 Interestingly, the protein A/G binding site on the Fc domain overlaps with the FcRn binding site, yet neither the variable or hinge domain of an IgG molecule influence the binding of an IgG to protein A/G protein.…”

Section: Resultsmentioning

confidence: 99%

Extending human IgG half-life using structure-guided design

Booth

Ramakrishnan

Narayan

et al. 2018

mAbs

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Engineering of antibodies for improved pharmacokinetics through enhanced binding to the neonatal Fc receptor (FcRn) has been demonstrated in transgenic mice, non-human primates and humans. Traditionally, such approaches have largely relied on random mutagenesis and display formats, which fail to address related critical attributes of the antibody, such as effector functions or biophysical stability. We have developed a structure- and network-based framework to interrogate the engagement of IgG with multiple Fc receptors (FcRn, C1q, TRIM21, FcγRI, FcγRIIa/b, FcγRIIIa) simultaneously. Using this framework, we identified features that govern Fc-FcRn interactions and identified multiple distinct pathways for enhancing FcRn binding in a pH-specific manner. Network analysis provided a novel lens to study the allosteric impact of half-life-enhancing Fc mutations on FcγR engagement, which occurs distal to the FcRn binding site. Applying these principles, we engineered a panel of unique Fc variants that enhance FcRn binding while maintaining robust biophysical properties and wild type-like binding to activating receptors. An antibody harboring representative Fc designs demonstrates a half-life improvement of > 9 fold in transgenic mice and > 3.5 fold in cynomolgus monkeys, and maintains robust effector functions such as antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity.

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Section: Resultsmentioning

confidence: 99%

Extending human IgG half-life using structure-guided design

Booth

Ramakrishnan

Narayan

et al. 2018

mAbs

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“…S1). A previous report suggested that this increase in apparent affinity may be due to charge driven interactions between the variable region of briakinumab and FcRn, 17 so we additionally assessed the affinity of F(ab')2 constructs. In concordance with the previous finding, briakinumab displayed binding to FcRn regardless of the buffer pH, while neither of the other antibodies displayed any appreciable binding ( Fig.…”

Section: Fcrn Affinity Measurement Via Biolayer Interferometrymentioning

confidence: 99%

“…[9][10][11][12][13][14] In addition to Fc engineering efforts, multiple studies have reported disparate PK parameters in antibodies with identical Fc regions but different variable regions. [15][16][17][18][19][20] In particular, clearance rates are increased in antibodies with polyreactive [21][22][23] or cross-reactive 24,25 profiles.…”

Section: Introductionmentioning

confidence: 99%

“…17,19,20 Affinity measurements in many of these experiments use surface plasmon resonance (SPR) or biolayer interferometry (BLI), conjugating FcRn to the sensor surface. As has been noted in previous studies, measurements for equivalent antibodies can vary significantly if great care is not taken, 14,26 and additionally the presence of a very small amount of aggregate can have a noticeable effect on apparent affinity measurement.…”

Section: Introductionmentioning

confidence: 99%

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Target-independent variable region mediated effects on antibody clearance can be FcRn independent

Kelly

Sun

et al. 2016

mAbs

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The importance of the neonatal Fc receptor (FcRn) in extending the serum half-life of monoclonal antibodies (mAbs) is well demonstrated, and has led to the development of multiple engineering approaches designed to alter Fc interactions with FcRn. Recent reports have additionally highlighted the effect of nonspecific interactions on antibody pharmacokinetics (PK), suggesting an FcRn-independent mechanism for mAb clearance. In this report we examine a case study of 2 anti-interleukin-12/23 antibodies, ustekinumab and briakinumab, which share the same target and Fc, but differ in variable region sequences. Ustekinumab displayed near baseline signal in a wide range of early stage developability assays for undesirable protein/protein interactions, while briakinumab showed significant propensity for self-and cross-interactions. This phenotypic difference correlates with faster clearance rates for briakinumab in both human FcRn transgenic and FcRn knockout mice. These findings support a dominant contribution for FcRn-independent clearance for antibodies with high nonspecificity, and highlight a key role for early stage developability screening to eliminate clones with such high nonspecific disposition PK.

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“…1–4 Recent studies have also shown that changes in the charge distribution on the protein surface could alter mAb pharmacokinetics (PK) by affecting FcRn-IgG dissociation. 5,6 Thus, the potential risk of deamidation at each site needs to be evaluated to ensure product stability.…”

Section: Introductionmentioning

confidence: 99%

Structure Based Prediction of Asparagine Deamidation Propensity in Monoclonal Antibodies

Yan

Huang

Lewis

et al. 2018

mAbs

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Identification of asparagine (Asn) sites that are prone to deamidation is critical for the development of therapeutic monoclonal antibodies (mAbs). Despite a common chemical degradation pathway, the rates of Asn deamidation can vary dramatically among different sites, and prediction of the sensitive deamidation sites is still challenging. In this study, characterization of Asn deamidation for five IgG1 and five IgG4 mAbs under both normal and stressed conditions revealed dramatic differences in the Asn deamidation rates. A comprehensive analysis of the deamidation sites indicated that the deamidation rate differences could be explained by differences in the local structure conformation, structure flexibility and solvent accessibility. A decision tree was developed to predict the deamidation propensity for all Asn sites in IgG mAbs based on the analysis of these three structural parameters. This decision tree will allow potential Asn deamidation hot spots to be identified early in development.

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Charge-mediated influence of the antibody variable domain on FcRn-dependent pharmacokinetics

Cited by 172 publications

References 46 publications

Extending human IgG half-life using structure-guided design

Extending human IgG half-life using structure-guided design

Target-independent variable region mediated effects on antibody clearance can be FcRn independent

Structure Based Prediction of Asparagine Deamidation Propensity in Monoclonal Antibodies

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