Forskning
Udskriv Udskriv
Switch language
Region Hovedstaden - en del af Københavns Universitetshospital
Udgivet

Diagnostic evaluation of a nanobody with picomolar affinity toward the protease RgpB from Porphyromonas gingivalis

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

DOI

  1. Nanograss sensor for selective detection of Pseudomonas aeruginosa by pyocyanin identification in airway samples

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  2. Variation in mitochondrial respiratory capacity and myosin heavy chain composition in repeated muscle biopsies

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  3. The best approach: homogenization or manual permeabilization of human skeletal muscle fibers for respirometry?

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  4. Small biomolecular scaffolds for improved biosensor performance

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  5. Micro method for determination of nonesterified fatty acid in whole blood obtained by fingertip puncture

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

  • Peter Durand Skottrup
  • Paul Leonard
  • Jakub Zbigniew Kaczmarek
  • Florian Veillard
  • Jan J. Enghild
  • Richard O'Kennedy
  • Aneta Sroka
  • Rasmus Prætorius Clausen
  • Jan Potempa
  • Erik Riise
Vis graf over relationer

Porphyromonas gingivalis is one of the major periodontitis-causing pathogens. P. gingivalis secretes a group of proteases termed gingipains, and in this study we have used the RgpB gingipain as a biomarker for P. gingivalis. We constructed a naive camel nanobody library and used phage display to select one nanobody toward RgpB with picomolar affinity. The nanobody was used in an inhibition assay for detection of RgpB in buffer as well as in saliva. The nanobody was highly specific for RgpB given that it did not bind to the homologous gingipain HRgpA. This indicated the presence of a binding epitope within the immunoglobulin-like domain of RgpB. A subtractive inhibition assay was used to demonstrate that the nanobody could bind native RgpB in the context of intact cells. The nanobody bound exclusively to the P. gingivalis membrane-bound RgpB isoform (mt-RgpB) and to secreted soluble RgpB. Further cross-reactivity studies with P. gingivalis gingipain deletion mutants showed that the nanobody could discriminate between native RgpB and native Kgp and RgpA in complex bacterial samples. This study demonstrates that RgpB can be used as a specific biomarker for P. gingivalis detection and that the presented nanobody-based assay could supplement existing methods for P. gingivalis detection.

OriginalsprogEngelsk
TidsskriftAnalytical Biochemistry
Vol/bind415
Udgave nummer2
Sider (fra-til)158-67
Antal sider10
ISSN0003-2697
DOI
StatusUdgivet - 15 aug. 2011

ID: 44863385