The Shark Strikes Twice: Hypervariable Loop 2 of Shark IgNAR Antibody Variable Domains and Its Potential to Function as an Autonomous Paratope.

Zielonka, Stefan; Empting, Martin; Könning, Doreen; Grzeschik, Julius; Krah, Simon; Becker, Stefan; Dickgießer, Stephan; Kolmar, Harald

dc.contributor.author	Zielonka, Stefan
dc.contributor.author	Empting, Martin
dc.contributor.author	Könning, Doreen
dc.contributor.author	Grzeschik, Julius
dc.contributor.author	Krah, Simon
dc.contributor.author	Becker, Stefan
dc.contributor.author	Dickgießer, Stephan
dc.contributor.author	Kolmar, Harald
dc.date.accessioned	2015-07-27T09:50:38Z	en
dc.date.available	2015-07-27T09:50:38Z	en
dc.date.issued	2015-08	en
dc.identifier.citation	The Shark Strikes Twice: Hypervariable Loop 2 of Shark IgNAR Antibody Variable Domains and Its Potential to Function as an Autonomous Paratope. 2015, 17 (4):386-92 Mar. Biotechnol.	en
dc.identifier.issn	1436-2236	en
dc.identifier.pmid	26003538	en
dc.identifier.doi	10.1007/s10126-015-9642-z	en
dc.identifier.uri	http://hdl.handle.net/10033/561055	en
dc.description.abstract	In this present study, we engineered hypervariable loop 2 (HV2) of the IgNAR variable domain in a way that it solely facilitates antigen binding, potentially functioning as an autonomous paratope. For this, the surface-exposed loop corresponding to HV2 was diversified and antigen-specific variable domain of IgNAR antibody (vNAR) molecules were isolated by library screening using yeast surface display (YSD) as platform technology. An epithelial cell adhesion molecule (EpCAM)-specific vNAR was used as starting material, and nine residues in HV2 were randomized. Target-specific clones comprising a new HV2-mediated paratope were isolated against cluster of differentiation 3ε (CD3ε) and human Fcγ while retaining high affinity for EpCAM. Essentially, we demonstrate that a new paratope comprising moderate affinities against a given target molecule can be engineered into the vNAR scaffold that acts independent of the original antigen-binding site, composed of complementarity-determining region 3 (CDR3) and CDR1.
dc.language.iso	en	en
dc.title	The Shark Strikes Twice: Hypervariable Loop 2 of Shark IgNAR Antibody Variable Domains and Its Potential to Function as an Autonomous Paratope.	en
dc.type	Article	en
dc.contributor.department	2Helmholtz-Institute for Pharmaceutical Research Saarland, Saarland University, Campus C2.3, 66123 Saarbrücken, Germany.	en
dc.identifier.journal	Marine biotechnology (New York, N.Y.)	en
refterms.dateFOA	2016-08-02T00:00:00Z
html.description.abstract	In this present study, we engineered hypervariable loop 2 (HV2) of the IgNAR variable domain in a way that it solely facilitates antigen binding, potentially functioning as an autonomous paratope. For this, the surface-exposed loop corresponding to HV2 was diversified and antigen-specific variable domain of IgNAR antibody (vNAR) molecules were isolated by library screening using yeast surface display (YSD) as platform technology. An epithelial cell adhesion molecule (EpCAM)-specific vNAR was used as starting material, and nine residues in HV2 were randomized. Target-specific clones comprising a new HV2-mediated paratope were isolated against cluster of differentiation 3ε (CD3ε) and human Fcγ while retaining high affinity for EpCAM. Essentially, we demonstrate that a new paratope comprising moderate affinities against a given target molecule can be engineered into the vNAR scaffold that acts independent of the original antigen-binding site, composed of complementarity-determining region 3 (CDR3) and CDR1.