Abstract
Immunophototherapy of cancer combines the specificity of a monoclonal antibody (MAb) to an overexpressed tumor marker with the phototoxic properties of the conjugated dye. To analyze the potential role of internalisation of the dye on photo-induced cytotoxicity, we compared two target antigens, carcinoembryonic antigen (CEA) that does not internalise and ErbB2 that does. Human ovarian carcinoma SKOv3 cells that express a high level of ErbB2 were transfected with the CEA cDNA. Using FACS analysis, the resulting cell line, SKOv3-CEA-1B9, demonstrated comparable levels of expression of the two target antigens. Aluminium tetrasulfophthalocyanine (AlPcS4) was covalently coupled to anti-CEA MAb 35A7, anti-ErbB2 MAb FSP77 and a non-specific MAb PX, via a five-carbon sulfonamide spacer chain (A1) at molar ratios ranging from 6 to 9 moles of AlPcS4 per mole of MAb. The 35A7-(AlPcS4A1)8 conjugate induced 68% growth inhibition of the SKOv3-CEA-1B9 cell line after a 20h incubation at 2.50μg/ml (based on AlPcS4A1 content) following light exposure. However, the FSP77-(AlPcS4A1)6 conjugate gave a 51% growth inhibition for an AlPcS4A1 concentration as low as 0.04μg/ml after the same incubation time and exposure to the same light dose. At a 1.25μg/ml AlPcS4A1 concentration, the FSP77-(AlPcS4A1)6 conjugate gave a 67% growth inhibition after an incubation time as short as 1h, reaching a 96% inhibition after an 8h incubation time. Using an unique cell line that expresses two different target antigens, we demonstrated a clear advantage of an internalising over a non-internalising MAb-dye conjugate in terms of phototoxic efficacy. in vivo evaluation of the photodynamic properties of the conjugates is in progress. © 2001 Cancer Research Campaign http://www.bjcancer.com
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16 November 2011
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Abbreviations
- MAb:
-
monoclonal antibody
- PBS:
-
phosphate-buffered saline
- EDC:
-
1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide hydrochloride
- sulfo-NHS:
-
N-hydroxysulfosuccinimide
- AlPcS4:
-
aluminium tetrasulfophthalocyanine
- CEA:
-
carcinoembryonic antigen
- Al:
-
five carbon spacer chain
- Px:
-
IgG1 purified from mouse myeloma P3-X63
- %ID/g:
-
percentage of injected dose per gram
- PS:
-
photosensitizer
- IPT:
-
immunophototherapy
- mTHPC:
-
metatetrahydroxyphenylchlorin
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Carcenac, M., Dorvillius, M., Garambois, V. et al. Internalisation enhances photo-induced cytotoxicity of monoclonal antibody-phthalocyanine conjugates. Br J Cancer 85, 1787–1793 (2001). https://doi.org/10.1054/bjoc.2001.2170
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DOI: https://doi.org/10.1054/bjoc.2001.2170
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