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Human monoclonal antibodies targeting carbonic anhydrase IX for the molecular imaging of hypoxic regions in solid tumours.

Ahlskog JK, Schliemann C, Mårlind J, Qureshi U, Ammar A, Pedley RB, Neri D - Br. J. Cancer (2009)

Bottom Line: These antibodies were able to stain CA IX ex vivo and to target the cognate antigen in vivo.In contrast, in a second animal model (SW1222), distinct staining patterns were observed for pimonidazole and CA IX targeting.We observed a complementary pattern of tumour regions targeted in vivo by the clinical-stage vascular-targeting antibody L19 and the anti-CA IX antibody A3, indicating that a homogenous pattern of in vivo tumour targeting could be achieved by a combination of the two antibodies.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry and Applied Biosciences, ETH Zürich, Wolfgang-Pauli-Strasse 10, Zurich CH-8093, Switzerland.

ABSTRACT

Background: Hypoxia, which is commonly observed in areas of primary tumours and of metastases, influences response to treatment. However, its characterisation has so far mainly been restricted to the ex vivo analysis of tumour sections using monoclonal antibodies specific to carbonic anhydrase IX (CA IX) or by pimonidazole staining, after the intravenous administration of this 2-nitroimidazole compound in experimental animal models.

Methods: In this study, we describe the generation of high-affinity human monoclonal antibodies (A3 and CC7) specific to human CA IX, using phage technology.

Results: These antibodies were able to stain CA IX ex vivo and to target the cognate antigen in vivo. In one of the two animal models of colorectal cancer studied (LS174T), CA IX imaging closely matched pimonidazole staining, with a preferential staining of tumour areas characterised by little vascularity and low perfusion. In contrast, in a second animal model (SW1222), distinct staining patterns were observed for pimonidazole and CA IX targeting. We observed a complementary pattern of tumour regions targeted in vivo by the clinical-stage vascular-targeting antibody L19 and the anti-CA IX antibody A3, indicating that a homogenous pattern of in vivo tumour targeting could be achieved by a combination of the two antibodies.

Conclusion: The new human anti-CA IX antibodies are expected to be non-immunogenic in patients with cancer and may serve as broadly applicable reagents for the non-invasive imaging of hypoxia and for pharmacodelivery applications.

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Biacore analysis of purified monomeric scFv preparations injected at different concentrations. (A) Binding of scFv(A3) and (B) scFv(CC7) to the extracellular CA domain of CA IX. Kinetic constants were calculated with the BIA evaluation 4.1 software.
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fig3: Biacore analysis of purified monomeric scFv preparations injected at different concentrations. (A) Binding of scFv(A3) and (B) scFv(CC7) to the extracellular CA domain of CA IX. Kinetic constants were calculated with the BIA evaluation 4.1 software.

Mentions: The monomeric fractions of the A3 and CC7 antibodies in recombinant scFv format were isolated by size-exclusion chromatography and analysed by real-time interaction analysis on a Biacore instrument, using a microsensor chip coated with the recombinant CA domain of CA IX. Figure 3 illustrates sensograms for the two antibodies, revealing a Kd dissociation constant of 2.4 nM for scFv(A3) [kon=9.1 × 105 s−1M−1; koff=2.2 × 10−3 s−1] and of 3.2 nM for scFv(CC7) [kon=4.3 × 105 s−1M−1; koff=1.4 × 10−3 s−1].


Human monoclonal antibodies targeting carbonic anhydrase IX for the molecular imaging of hypoxic regions in solid tumours.

Ahlskog JK, Schliemann C, Mårlind J, Qureshi U, Ammar A, Pedley RB, Neri D - Br. J. Cancer (2009)

Biacore analysis of purified monomeric scFv preparations injected at different concentrations. (A) Binding of scFv(A3) and (B) scFv(CC7) to the extracellular CA domain of CA IX. Kinetic constants were calculated with the BIA evaluation 4.1 software.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2736829&req=5

fig3: Biacore analysis of purified monomeric scFv preparations injected at different concentrations. (A) Binding of scFv(A3) and (B) scFv(CC7) to the extracellular CA domain of CA IX. Kinetic constants were calculated with the BIA evaluation 4.1 software.
Mentions: The monomeric fractions of the A3 and CC7 antibodies in recombinant scFv format were isolated by size-exclusion chromatography and analysed by real-time interaction analysis on a Biacore instrument, using a microsensor chip coated with the recombinant CA domain of CA IX. Figure 3 illustrates sensograms for the two antibodies, revealing a Kd dissociation constant of 2.4 nM for scFv(A3) [kon=9.1 × 105 s−1M−1; koff=2.2 × 10−3 s−1] and of 3.2 nM for scFv(CC7) [kon=4.3 × 105 s−1M−1; koff=1.4 × 10−3 s−1].

Bottom Line: These antibodies were able to stain CA IX ex vivo and to target the cognate antigen in vivo.In contrast, in a second animal model (SW1222), distinct staining patterns were observed for pimonidazole and CA IX targeting.We observed a complementary pattern of tumour regions targeted in vivo by the clinical-stage vascular-targeting antibody L19 and the anti-CA IX antibody A3, indicating that a homogenous pattern of in vivo tumour targeting could be achieved by a combination of the two antibodies.

View Article: PubMed Central - PubMed

Affiliation: Department of Chemistry and Applied Biosciences, ETH Zürich, Wolfgang-Pauli-Strasse 10, Zurich CH-8093, Switzerland.

ABSTRACT

Background: Hypoxia, which is commonly observed in areas of primary tumours and of metastases, influences response to treatment. However, its characterisation has so far mainly been restricted to the ex vivo analysis of tumour sections using monoclonal antibodies specific to carbonic anhydrase IX (CA IX) or by pimonidazole staining, after the intravenous administration of this 2-nitroimidazole compound in experimental animal models.

Methods: In this study, we describe the generation of high-affinity human monoclonal antibodies (A3 and CC7) specific to human CA IX, using phage technology.

Results: These antibodies were able to stain CA IX ex vivo and to target the cognate antigen in vivo. In one of the two animal models of colorectal cancer studied (LS174T), CA IX imaging closely matched pimonidazole staining, with a preferential staining of tumour areas characterised by little vascularity and low perfusion. In contrast, in a second animal model (SW1222), distinct staining patterns were observed for pimonidazole and CA IX targeting. We observed a complementary pattern of tumour regions targeted in vivo by the clinical-stage vascular-targeting antibody L19 and the anti-CA IX antibody A3, indicating that a homogenous pattern of in vivo tumour targeting could be achieved by a combination of the two antibodies.

Conclusion: The new human anti-CA IX antibodies are expected to be non-immunogenic in patients with cancer and may serve as broadly applicable reagents for the non-invasive imaging of hypoxia and for pharmacodelivery applications.

Show MeSH
Related in: MedlinePlus