Structural analysis of Fc/FcγR complexes: a blueprint for antibody design

Caaveiro, José M. M.; Kiyoshi, Masato; Tsumoto, Kouhei

doi:10.1111/imr.12365

Cited by 67 publications

(67 citation statements)

References 159 publications

(212 reference statements)

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“…Measuring the FcγR-activating capabilities of antiviral IgG augments definition of immune correlates of protection against infections and/or infection-induced disease progression. Three different types of Fcγ receptors are displayed on the cell surface of human leukocytes: FcγRI (CD64), FcγRII (types A, B, and C, collectively known as CD32), and FcγRIII (types A and B, collectively known as CD16) 14 . Binding affinity of human IgG Fc to a corresponding FcγR is dictated by both the IgG-Fc subclass (IgG1, IgG2, IgG3 and IgG4) and changes in a single N-linked glycan located in the CH2 domain of the IgG Fc 15–18 .…”

Section: Introductionmentioning

confidence: 99%

Sudan ebolavirus long recovered survivors produce GP-specific Abs that are of the IgG1 subclass and preferentially bind FcγRI

Radinsky

Edri

Brusilovsky

et al. 2017

Sci Rep

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Ebolavirus is a highly lethal pathogen, causing a severe hemorrhagic disease with a high fatality rate. To better understand immune correlates of protection by virus specific IgG, we investigated the evolution of the Fcγ receptors (FcγRs)-activating capabilities of antiviral IgG in serum samples of long recovered survivors. To this end, longitudinal serum samples from survivors of Sudan ebolavirus (SUDV) infection, studied over years, were examined for the presence of Ebola-GP specific IgG subclasses, and for their binding to FcγRs. We developed a cell-based reporter system to quantitate pathogen-specific antibody binding to FcγRIIIA, FcγRIIA, FcγRIIB and FcγRI. With this system, we demonstrate that anti-GP-specific stimulation of the FcγRI reporter by survivors’ sera was substantially high one year after acute infection, with a slight reduction in activity over a decade post infection. We further demonstrate that GP-specific IgG1 is by far the seroprevalent subclass that retained and even enhanced its presence in the sera, over ten years post infection; the prevalence of other GP-specific IgG subclasses was considerably reduced over time. In accordance, GP-specific FcγRI reporter response and GP-specific total IgG1 subclass correlated in the studied group of Ebola survivors. These observations are important for further informing Ebola vaccine and therapeutic development.

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Section: Introductionmentioning

confidence: 99%

Sudan ebolavirus long recovered survivors produce GP-specific Abs that are of the IgG1 subclass and preferentially bind FcγRI

Radinsky

Edri

Brusilovsky

et al. 2017

Sci Rep

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“…In addition, AA residues participating in the binding energy were shown to be mainly aromatic residues, as well as Gly or Ser . In several works, remarkable conformational changes in the structure of Abs and antigen during the formation of their complexes were observed …”

Section: Discussionmentioning

confidence: 99%

How human IgGs against myelin basic protein (MBP) recognize oligopeptides and MBP

2017

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Myelin basic protein (MBP) is a major protein of myelin-proteolipid shell of axons, and it plays an important role in pathogenesis of multiple sclerosis. In the literature, there are no data on how antibodies recognize different protein antigens including MBP. A stepwise increase in ligand complexity was used to estimate the relative contributions of virtually every amino acid residue (AA) of a specific 12-mer LSRFSWGAEGQK oligopeptide corresponding to immunodominant sequence of MBP to the light chains and to intact anti-MBP IgGs from sera of patients with multiple sclerosis. It was shown that the minimal ligands of the light chains of IgGs are many different free AAs (K = 0.51-0.016 M), and each free AA interacts with the specific subsite of the light chain intended for recognition of this AA in specific LSRFSW oligopeptide. A gradual transition from Leu to LSRFSWGAEGQK leads to an increase in the affinity from 10 to 2.3 × 10 M because of additive interactions of the light chain with 6 AAs of this oligopeptide and then the affinity reaches plateau. The contributions of 6 various AAs to the affinity of the oligopeptide are different (K , M): 0.71 (S), 0.44 (R), 0.14 (F), 0.17 (S), and 0.62 (W). Affinity of nonspecific oligopeptides to the light chains of IgGs is significantly lower. Intact MBP interacts with both light and heavy chains of IgGs demonstrating 192-fold higher affinity than the specific oligopeptide. It is a first quantitative analysis of the mechanism of proteins recognition by antibodies. The thermodynamic model was constructed to describe the interactions of IgGs with MBP. The data obtained can be very useful for understanding how antibodies against many different proteins can recognize these proteins.

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“…Fusions containing this truncated Fc region are unlikely to retain effector functions because Fcg receptor binding is known to involve the N-terminal region of the CH2 domain. 20 This is generally unimportant for in vitro applications; however, if retention of Fcg receptor binding were required, it may be possible to restore this function by adding back some or all of the N-terminal CH2 residues.…”

Section: Discussionmentioning

confidence: 99%

A hingeless Fc fusion system for site-specific cleavage by IdeS

Novarra¹,

Grinberg²,

Rickert³

et al. 2016

mAbs

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Fusion of proteins to the Fc region of IgG is widely used to express cellular receptors and other extracellular proteins, but cleavage of the fusion partner is sometimes required for downstream applications. Immunoglobulin G-degrading enzyme of Streptococcus pyogenes (IdeS) is a protease with exquisite specificity for human IgG, and it can also cleave Fc-fusion proteins at a single site in the Nterminal region of the CH2 domain. However, the site of IdeS cleavage results in the disulfide-linked hinge region partitioning with the released protein, complicating downstream usage of the cleaved product. To tailor the Fc fragment for release of partner proteins by IdeS treatment, we investigated the effect of deleting regions of IgG-derived sequence that are upstream of the cleavage site. Elimination of the IgGderived hinge sequence along with several residues of the CH2 domain had negligible effects on expression and purity of the fusion protein, while retaining efficient processing by IdeS. An optimal Fc fragment comprising residues 235-447 of the human IgG1 heavy chain sufficed for efficient production of fusion proteins and minimized the amount of residual Ig-derived sequence on the cleavage product following IdeS treatment. Pairing of this truncated Fc fragment with IdeS cleavage enables highly specific cleavage of Fc-fusion proteins, thus eliminating the need to engineer extraneous cleavage sequences. This system should be helpful for producing Fc-fusion proteins requiring downstream cleavage, particularly those that are sensitive to internal miscleavage if treated with alternative proteases.Abbreviations: EPOR, erythropoietin receptor; Fc, fragment crystallizable; IdeS, Immunoglobulin G-degrading enzyme of Streptococcus pyogenes; mAb, monoclonal antibody; PBS, phosphate-buffered saline; SEC, size exclusion chromatography; scFv, single chain variable fragment; TRAILR2, TNF-related apoptosis inducing ligand receptor 2

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Structural analysis of Fc/FcγR complexes: a blueprint for antibody design

Cited by 67 publications

References 159 publications

Sudan ebolavirus long recovered survivors produce GP-specific Abs that are of the IgG1 subclass and preferentially bind FcγRI

Sudan ebolavirus long recovered survivors produce GP-specific Abs that are of the IgG1 subclass and preferentially bind FcγRI

How human IgGs against myelin basic protein (MBP) recognize oligopeptides and MBP

A hingeless Fc fusion system for site-specific cleavage by IdeS

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