Construction and characterization of the chimeric monoclonal antibody E48 for therapy of head and neck cancer.
Data from an ongoing clinical radioimmunoscintigraphy trial indicate that 99mTc-labeled monoclonal antibody (mAb) E48 is highly capable of selectively targeting squamous cell carcinoma of the head and neck (HNSCC). The percentage of the injected dose per gram of tumor tissue was found to be high, rendering mAb E48 a promising candidate mAb for therapeutic purposes. We now describe the construction of a chimeric (mouse/human) mAb E48 by recombinant DNA technology. The genes encoding the variable domains of the heavy and light chain were cloned and ligated into expression vectors containing the human gamma 1 heavy-chain gene and the human kappa light-chain gene respectively. Biological properties of the resulting chimeric mAb E48 were compared to the murine form in vitro and in vivo. The reactivities of chimeric (c)mAb and murine (m)mAb E48 with HNSCC, as assessed by immunohistochemical staining as well as immuno-blotting were shown to be similar. The affinity constant appeared to be 0.9 x 10(10) M-1 and 1.6 x 10(10) M-1 for the mmAb and cmAb respectively. The biodistribution of both antibodies was tested by simultaneous injection into nude mice bearing human HNSCC xenografts. cmAb E48 was found to be cleared more rapidly from the blood than mmAb E48, resulting in a 30% lower tumor uptake but similar tumor to non-tumor ratios, 3 days after injection. Moreover, it was shown that cmAb E48 is highly capable of lysing HNSCC targets in ADCC assays in vitro, whereas the mmAb appeared to be almost inactive. These data indicate that cmAb E48 has potential as a targeting agent for the eradication of HNSCC in man.
Brakenhoff
R H
RH
Department of Otorhinolaryngology/Head and Neck Surgery, Free University Hospital, Amsterdam, The Netherlands.
van Gog
F B
FB
Looney
J E
JE
van Walsum
M
M
Snow
G B
GB
van Dongen
G A
GA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics*; D000920:Antibody-Dependent Cell Cytotoxicity / Q000276:immunology; D001483:Base Sequence; D002294:Carcinoma, Squamous Cell / Q000276:immunology / Q000628:therapy*; D003001:Cloning, Molecular / Q000379:methods; D006258:Head and Neck Neoplasms / Q000276:immunology / Q000628:therapy*; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007167:Immunotherapy; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*
D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins