Novel anti-human Axl monoclonal antibodies for improved patient biomarker studies

  • Lavina Ahmed BerGenBio AS and Center for Cancer Biomarkers CCBIO, Department of Clinical Medicine, University of Bergen, Norway
  • Sergej M Kiprijanov BerGenBio AS
  • Hawa nalwoga Center for Cancer Biomarkers CCBIO, Department of Clinical Medicine, University of Bergen, Norway
  • Katarzyna Wnuk-Lipinska BerGenBio AS
  • Hallvard Haugen BerGenBio AS
  • James B Lorens BerGenBio AS and University of Bergen, Norway
  • Lars A Akslen Centre for Cancer Biomarkers, University of Bergen, Norway and Department of Pathology, Haukeland University Hospital, Bergen, Norway
  • David Robert Micklem BerGenBio AS http://orcid.org/0000-0003-2756-2591

Abstract

Background: Axl receptor tyrosine kinase has been associated with poor clinical outcome in a wide variety of malignancies including breast, lung, prostate, pancreatic, brain and myeloid cancers. Axl is up-regulated during tumour epithelial-to-mesenchymal transition (EMT) and is associated with metastasis, immune evasion and drug resistance.
Methods: The aim of this study was to develop improved assays for detection of Axl protein in patient samples. We developed a panel of mouse monoclonal antibodies directed against the extracellular domain of human Axl, from mice immunized with a recombinant antigen comprising the extracellular domains of human Axl fused to human IgG1 Fc domain. Anti-Axl monoclonal antibodies (MAbs) were assessed for sensitivity, specificity and utility using surface plasmon resonance (SPR) analysis, sandwich enzyme-linked immunosorbent assay (ELISA), Western blot analysis and immunohistochemistry (IHC) staining.
Results: One of the anti-human Axl MAb,1H12, performed particularly well, and SPR analysis confirmed both high affinity (50 pM) and high selectivity for Axl versus other Axl-family receptors. In IHC applications on selected human carcinomas and cancer cell line pellets, MAb 1H12 showed improved sensitivity and reduced background staining compared to other members of the MAb panel. A second MAb 5F11 showed very high affinity (5.8 pM) at a non-overlapping epitope and was selected as a partner for 1H12 in sandwich ELISA, producing a highly sensitive and specific assay.
Conclusion: MAbs 1H12 and 5F11 represent new anti-human Axl monoclonal antibodies for improved patient biomarker studies.

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Author Biographies

Lavina Ahmed, BerGenBio AS and Center for Cancer Biomarkers CCBIO, Department of Clinical Medicine, University of Bergen, Norway
Industrial PhD student BerGenBio AS/University of Bergen
Sergej M Kiprijanov, BerGenBio AS
Director of Biologics Research and Development, BerGenBio AS
Hawa nalwoga, Center for Cancer Biomarkers CCBIO, Department of Clinical Medicine, University of Bergen, Norway
Postdoctoral fellow, Department of Clinical Medicine
Katarzyna Wnuk-Lipinska, BerGenBio AS

Postdoctoral researcher, BerGenBio AS

When this work was performed, KWL was an Industrial PhD student with BerGenBio/Department of Biomedicine, University of Bergen

Hallvard Haugen, BerGenBio AS

BerGenBio AS

When this work was performed, HH was an Industrial PhD student with BerGenBio/Department of Biomedicine, University of Bergen

James B Lorens, BerGenBio AS and University of Bergen, Norway

Chief Scientific Officer, BerGenBio AS

Professor, Department of Biomedicine and Centre for Cancer Biomarkers, University of Bergen

Lars A Akslen, Centre for Cancer Biomarkers, University of Bergen, Norway and Department of Pathology, Haukeland University Hospital, Bergen, Norway

Director, Centre for Cancer Biomarkers CCBIO, University of Bergen, Norway

Professor, Department of Pathology, Haukeland University Hospital, Bergen, Norway

David Robert Micklem, BerGenBio AS
Director of Diagnostics and Biomarkers

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Published
2016-04-20
How to Cite
AHMED, Lavina et al. Novel anti-human Axl monoclonal antibodies for improved patient biomarker studies. Diagnostic Pathology, [S.l.], v. 2, n. 1, apr. 2016. ISSN 2364-4893. Available at: <http://www.diagnosticpathology.eu/content/index.php/dpath/article/view/104>. Date accessed: 26 nov. 2020. doi: https://doi.org/10.17629/www.diagnosticpathology.eu-2016-2:104.
Section
Research

Keywords

Monoclonal antibodies; cancer; EMT; diagnostic biomarkers; Axl; ELISA; IHC; Western blot