A naturally occurring NAR variable domain binds the Kgp protease from <i>Porphyromonas gingivalis</i>

Nuttall, Stewart D.; Krishnan, Usha V.; Doughty, Larissa; Nathanielsz, Anne; Ally, Nafisa; Pike, Robert N.; Hudson, Peter J.; Kortt, Alexander A.; Irving, Robert A.

doi:10.1016/s0014-5793(02)02506-1

Cited by 61 publications

(54 citation statements)

References 21 publications

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“…8, which is published as supporting information on the PNAS web site) and in other recombinant V NAR s proteins (ref. 9 and data not shown).…”

Section: Resultsmentioning

confidence: 76%

“…However, recently, a distinctly unconventional antibody isotype was identified in the serum of nurse sharks (Ginglymostoma cirratum) and wobbegong sharks (Orectolobus maculatus): the Ig new antigen receptors (IgNARs) (6,7). Current evidence implicates IgNARs as true molecules of the immune armory and as the most probable agent of the shark antigen-driven affinity-maturation antibody response (8)(9)(10).…”

mentioning

confidence: 99%

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Structural evidence for evolution of shark Ig new antigen receptor variable domain antibodies from a cell-surface receptor

Streltsov

Varghese²,

Carmichael³

et al. 2004

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

120

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The Ig new antigen receptors (IgNARs) are single-domain antibodies found in the serum of sharks. Here, we report 2.2-and 2.8-Å structures of the type 2 IgNAR variable domains 12Y-1 and 12Y-2. Structural features include, first, an Ig superfamily topology transitional between cell adhesion molecules, antibodies, and T cell receptors; and, second, a vestigial complementarity-determining region 2 at the ''bottom'' of the molecule, apparently discontinuous from the antigen-binding paratope and similar to that observed in cell adhesion molecules. Thus, we suggest that IgNARs originated as cell-surface adhesion molecules coopted to the immune repertoire and represent an evolutionary lineage independent of variable heavy chain͞variable light chain type antibodies. Additionally, both 12Y-1 and 12Y-2 form unique crystallographic dimers, predominantly mediated by main-chain framework interactions, which represent a possible model for primordial cell-based interactions. Unusually, the 12Y-2 complementarity-determining region 3 also adopts an extended ␤-hairpin structure, suggesting a distinct selective advantage in accessing cryptic antigenic epitopes.

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“…8, which is published as supporting information on the PNAS web site) and in other recombinant V NAR s proteins (ref. 9 and data not shown).…”

Section: Resultsmentioning

confidence: 76%

mentioning

confidence: 99%

Structural evidence for evolution of shark Ig new antigen receptor variable domain antibodies from a cell-surface receptor

Streltsov

Varghese²,

Carmichael³

et al. 2004

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

120

View full text Add to dashboard Cite

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“…43 It has been well established that IgNAR is amenable to phage display-immunized, naïve and semi-synthetic IgNAR phage display libraries have been reported in the literature. 42,[45][46][47][48][49][50] Here too, it proved an effective platform for the iterative enrichment and isolation of antigen specific binders. As serum albumin binds in a pH-dependent manner to FcRn, it was critical to incorporate this parameter in the post-selection characterization of lead clones in addition to cross-reactivity with MSA to fit with our initial murine PK model.…”

Section: Construction and Screening Of The Immunized Vnar Phage Librarymentioning

confidence: 99%

Improving the pharmacokinetic properties of biologics by fusion to an anti-HSA shark VNAR domain

Müller

Saunders

Grace

et al. 2012

mAbs

105

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“…Recombinant proteins were expressed in the bacterial periplasm as described previously [27,28]. Proteins were purified from the periplasm by the method of Minsky [29] and used either as crude fractions or purified further by affinity chromatography using ANTI-FLAG M2 Affinity gel resin (2-10 × 1 cm) (Sigma, St. Louis, MO, USA).…”

Section: Soluble Expression Of V Nar Constructs From Expression Vectomentioning

confidence: 99%

Shark Variable New Antigen Receptor (VNAR) Single Domain Antibody Fragments: Stability and Diagnostic Applications

et al. 2013

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Abstract:The single variable new antigen receptor domain antibody fragments (V NAR s) derived from shark immunoglobulin new antigen receptor antibodies (IgNARs) represent some of the smallest known immunoglobulin-based protein scaffolds. As single domains, they demonstrate favorable size and cryptic epitope recognition properties, making them attractive in diagnosis and therapy of numerous disease states. Here, we examine the stability of V NAR domains with a focus on a family of V NAR s specific for apical membrane antigen 1 (AMA-1) from Plasmodium falciparum. The V NAR s are compared to traditional monoclonal antibodies (mAbs) in liquid, lyophilized and immobilized nitrocellulose formats. When maintained in various formats at 45 °C, V NAR s have improved stability compared to mAbs for periods of up to four weeks. Using circular dichroism spectroscopy we demonstrate that V NAR domains are able to refold following heating to 80 °C. We also demonstrate that V NAR domains are stable during incubation under potential in vivo conditions such as stomach acid, but not to the protease rich environment of murine stomach scrapings. Taken together, our results demonstrate the suitability of shark V NAR domains for various diagnostic platforms and related applications.

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A naturally occurring NAR variable domain binds the Kgp protease from Porphyromonas gingivalis

Cited by 61 publications

References 21 publications

Structural evidence for evolution of shark Ig new antigen receptor variable domain antibodies from a cell-surface receptor

Structural evidence for evolution of shark Ig new antigen receptor variable domain antibodies from a cell-surface receptor

Improving the pharmacokinetic properties of biologics by fusion to an anti-HSA shark VNAR domain

Shark Variable New Antigen Receptor (VNAR) Single Domain Antibody Fragments: Stability and Diagnostic Applications

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