Complex of a Protective Antibody with Its Ebola Virus GP Peptide Epitope: Unusual Features of a Vλx Light Chain

Lee, Jeffrey E.; Kuehne, Ana I.; Abelson, Dafna M.; Fusco, Marnie L.; Hart, Mary Kate; Saphire, Erica Ollmann

doi:10.1016/j.jmb.2007.10.017

Cited by 52 publications

(39 citation statements)

References 45 publications

(9 reference statements)

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“…Alternatively, non-Fc amino acids may have a conformational affect on the Fc region. h-13F6 contains a rare Vλx light-chain variable region and the crystal structure for the murine parental mAb shows that the three light-chain complementarity-determining regions adopt unusual conformations distinct from Vκ and other Vλ light chains (19). Also noteworthy is the relatively high affinity of h-13F6 agly for FcγRIII.…”

Section: Discussionmentioning

confidence: 99%

Enhanced potency of a fucose-free monoclonal antibody being developed as an Ebola virus immunoprotectant

Zeitlin

Pettitt

Scully

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

205

195

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No countermeasures currently exist for the prevention or treatment of the severe sequelae of Filovirus (such as Ebola virus; EBOV) infection. To overcome this limitation in our biodefense preparedness, we have designed monoclonal antibodies (mAbs) which could be used in humans as immunoprotectants for EBOV, starting with a murine mAb (13F6) that recognizes the heavily glycosylated mucin-like domain of the virion-attached glycoprotein (GP). Point mutations were introduced into the variable region of the murine mAb to remove predicted human T-cell epitopes, and the variable regions joined to human constant regions to generate a mAb (h-13F6) appropriate for development for human use. We have evaluated the efficacy of three variants of h-13F6 carrying different glycosylation patterns in a lethal mouse EBOV challenge model. The pattern of glycosylation of the various mAbs was found to correlate to level of protection, with aglycosylated h-13F6 providing the least potent efficacy (ED 50 = 33 μg). A version with typical heterogenous mammalian glycoforms (ED 50 = 11 μg) had similar potency to the original murine mAb. However, h-13F6 carrying complex N-glycosylation lacking core fucose exhibited superior potency (ED 50 = 3 μg). Binding studies using Fcγ receptors revealed enhanced binding of nonfucosylated h-13F6 to mouse and human FcγRIII. Together the results indicate the presence of Fc N-glycans enhances the protective efficacy of h-13F6, and that mAbs manufactured with uniform glycosylation and a higher potency glycoform offer promise as biodefense therapeutics.passive immunization | antibody glycosylation | antibody-dependent cellular cytotoxicity | antiviral

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Section: Discussionmentioning

confidence: 99%

Enhanced potency of a fucose-free monoclonal antibody being developed as an Ebola virus immunoprotectant

Zeitlin

Pettitt

Scully

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

205

195

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“…Thus, even though we have structural information on the lower core of GP, there is no structural information on the mucin-like domain, save one structure of an antibody bound to a separate mucin-like domain linear epitopes. This crystal structure, of MAb 13F6-1-2, a representative member of competition group I, in complex with its peptide linear epitope (GP residues 401 to 417), has provided insight into its unique structural mechanism of neutralization (23). 13F6-1-2 contains an extremely rare Vx light chain that is represented by only 0.5% of mouse antibody sequences.…”

mentioning

confidence: 99%

“…13F6-1-2 contains an extremely rare Vx light chain that is represented by only 0.5% of mouse antibody sequences. The use of this very rare light chain confers noncanonical structures to all three CDRs and may consequently result in the unusual mode of binding of the antibody (23). The GP 401-417 peptide, representing the epitope of 13F6-1-2, adopts a linear, extended conformation and binds into a shallow groove in the 13F6-1-2 combining site that extends diagonally across the antibody combining site.…”

mentioning

confidence: 99%

“…The GP 401-417 peptide, representing the epitope of 13F6-1-2, adopts a linear, extended conformation and binds into a shallow groove in the 13F6-1-2 combining site that extends diagonally across the antibody combining site. Two particular amino acids of GP 401-417 , Gly P406 and Arg P409, form extensive contacts (eight in all) with 13F6 and are critical for antibody recognition (23).…”

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confidence: 99%

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Structure of an Antibody in Complex with Its Mucin Domain Linear Epitope That Is Protective against Ebola Virus

Olal

Kuehne

Bale

et al. 2012

J Virol

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Antibody 14G7 is protective against lethal Ebola virus challenge and recognizes a distinct linear epitope in the prominent mucinlike domain of the Ebola virus glycoprotein GP. The structure of 14G7 in complex with its linear peptide epitope has now been determined to 2.8 Å. The structure shows that this GP sequence forms a tandem ␤-hairpin structure that binds deeply into a cleft in the antibody-combining site. A key threonine at the apex of one turn is critical for antibody interaction and is conserved among all Ebola viruses. This work provides further insight into the mechanism of protection by antibodies that target the protruding, highly accessible mucin-like domain of Ebola virus and the structural framework for understanding and characterizing candidate immunotherapeutics.

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“…To visualize potential roles of Vλx junctional diversity in Ag binding, we compared the structures of three Abs using Vλx that have been determined by X-ray diffraction (17,28,29). In each case, the structure of the unbound Abs and of the Ab-Ag complexes could be examined.…”

Section: Resultsmentioning

confidence: 99%

Antibodies that bind complex glycosaminoglycans accumulate in the Golgi

Radic

Weigert

Khan

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Light (L) chains that edit anti-DNA heavy (H) chains rescue B-cell development by suppressing DNA binding. However, exceptional editor L chains allow B cells to reach splenic compartments even though their B-cell receptors remain autoreactive. Such incompletely edited B cells express multireactive antibodies that accumulate in the Golgi and are released as insoluble, amyloid-like immune complexes. Here, we examine examples of incomplete editing from the analysis of variable to joining (VJ) gene junction of the variable (Vλx) editor L chain. When paired with the anti-DNA heavy chain, V H 56R, the Vλx variants yield antibodies with differing specificities, including glycosaminoglycan reactivity. Our results implicate these specificities in the evasion of receptor editing through intracellular sequestration of IgM and the release of insoluble IgM complexes. Our findings can be extrapolated to human L chains and have implications for understanding a latent component of the Ig repertoire that could exert pathogenic and protective functions.

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Complex of a Protective Antibody with Its Ebola Virus GP Peptide Epitope: Unusual Features of a Vλx Light Chain

Cited by 52 publications

References 45 publications

Enhanced potency of a fucose-free monoclonal antibody being developed as an Ebola virus immunoprotectant

Enhanced potency of a fucose-free monoclonal antibody being developed as an Ebola virus immunoprotectant

Structure of an Antibody in Complex with Its Mucin Domain Linear Epitope That Is Protective against Ebola Virus

Antibodies that bind complex glycosaminoglycans accumulate in the Golgi

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