Structural characterization of a human Fc fragment engineered for extended serum half-life

Oganesyan, Vaheh; Damschroder, Melissa; Woods, R. Claude; Cook, Kimberly E.; Wu, Herren; Dall’Acqua, William F.

doi:10.1016/j.molimm.2009.01.026

Cited by 81 publications

(59 citation statements)

References 28 publications

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“…Expression and Purification of Fc-YTE-A human Fc containing the M252Y/S254T/T256E ("YTE") mutations was expressed in HEK 293 cells and purified as previously described (18). Purified Fc-YTE was further dialyzed overnight at 4°C against 50 mM NaOAc, pH 5.2, 100 mM NaCl and concentrated to ϳ4 mg/ml.…”

Section: Methodsmentioning

confidence: 99%

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Structural Insights into Neonatal Fc Receptor-based Recycling Mechanisms

Oganesyan¹,

Damschroder²,

Cook³

et al. 2014

Journal of Biological Chemistry

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213

298

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Section: Methodsmentioning

confidence: 99%

“…Determination of the HSA/FcRn/Fc-YTE Three-dimensional Structure-HSA/FcRn (this study) and Fc-YTE (PDB entry 3FJT) (18) were used as models. The unit cell with parameters a ϭ b ϭ 153.2, c ϭ 146.0 Å contained one FcRn, one HSA, and one Fc-YTE polypeptide (half-Fc).…”

Section: Generation and Expression Of Human Igg And Igg-yte Mutants-hmentioning

confidence: 99%

Structural Insights into Neonatal Fc Receptor-based Recycling Mechanisms

Oganesyan¹,

Damschroder²,

Cook³

et al. 2014

Journal of Biological Chemistry

Self Cite

213

298

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“…33 Among many reported mutations, the triple mutations M252Y-S254T-T256E (referred to hereafter as YTE), resulted in increased binding to both cynomolgus monkey and human FcRn at pH 6, while preserving no binding at pH 7.4. 34,35 To test whether the Bs4Ab platform allows Fc modifications similarly to IgG1, we prepared a Bs4Ab-VA with the YTE, TM and 3M mutations and performed a comparative analysis using the anti-VEGF antibody modified with the same Fc mutations. Proteins were transiently expressed using HEK293 cells and expression levels after 10 d of culture were 130 mg/mL, 125 mg/mL and 85 mg/mL for Bs4Ab-VA-YTE, Bs4Ab-VA-TM and Bs4Ab-3M, respectively.…”

Section: Examples Of Antibodies Derived From the Bs4ab Technologymentioning

confidence: 99%

Insertion of scFv into the hinge domain of full-length IgG1 monoclonal antibody results in tetravalent bispecific molecule with robust properties

Bezabeh¹,

Fleming²,

Fazenbaker³

et al. 2016

mAbs

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By simultaneous binding two disease mediators, bispecific antibodies offer the opportunity to broaden the utility of antibody-based therapies. Herein, we describe the design and characterization of Bs4Ab, an innovative and generic bispecific tetravalent antibody platform. The Bs4Ab format comprises a full-length IgG1 monoclonal antibody with a scFv inserted into the hinge domain. The Bs4Ab design demonstrates robust manufacturability as evidenced by MEDI3902, which is currently in clinical development. To further demonstrate the applicability of the Bs4Ab technology, we describe the molecular engineering, biochemical, biophysical, and in vivo characterization of a bispecific tetravalent Bs4Ab that, by simultaneously binding vascular endothelial growth factor and angiopoietin-2, inhibits their function. We also demonstrate that the Bs4Ab platform allows Fc-engineering similar to that achieved with IgG1 antibodies, such as mutations to extend half-life or modulate effector functions.

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“…In a study aimed to demonstrate positive correlation between FcRn binding affinity and effector anti-tumor activity, involving hydrogen bonds. 49 In a different human IgG1 with mutations to enhance ADCC activity by increased binding to FcγRIIIA (etaracizumab, anti-human α v β 3 integrin complex, MedImmune), introduction of YTE residues resulted in >100-fold reduction in the ADCC activity. This negative effect was reversed with additional mutations that consisted of S239D/A330L/I332E and reconstitution of the ADCC activity to 10-fold that of the original MEDI-522.…”

Section: Aaa From a Previous Comprehensive Screen Of Fc Mutations Inmentioning

confidence: 99%