Generation and characterization of a novel single-gene-encoded single-chain immunoglobulin molecule with antigen binding activity and effector functions

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Title
Generation and characterization of a novel single-gene-encoded single-chain immunoglobulin molecule with antigen binding activity and effector functions
Author(s)
Hyun Sil Lee; Liming Shu; Roberto De Pascalis; Mariateresa Giuliano; Mingzhu Zhu; Eduardo A Padlan; Patricia Horan Hand; Jeffrey Schlom; Hyo Jeong Hong; S V S Kashmiri
Bibliographic Citation
Molecular Immunology, vol. 36, pp. 61-71
Publication Year
1999
Abstract
Monoclonal antibody (MAb) CC49 is a murine IgG1 that reacts with tumor- associated glycoprotein (TAG)-72, a pancarcinoma antigen. Clinical trials using radiolabeled CC49 for diagnostic imaging have demonstrated specific localization of more than 90% of carcinomas. The feasibility of adopting in vivo gene inoculation methods for antibody-based immunotherapy requires introduction and expression of two genes, encoding immunoglobulin (Ig) heavy and light chains, in a single cell to generate a functional antibody. To circumvent the problems inherent in this approach, we have constructed a single-gene encoding a single-chain immunoglobulin (SCIg) that, unlike previously developed SCIgs, contains all IgG domains. To construct the novel SCIg, the carboxyl end of the constant region of the chimeric (c) CC49 kappa chain is joined, via a 30 residue Gly-Ser linker peptide, to the amino terminus of the CC49 heavy chain. To our knowledge, neither a linker peptide this long nor a linkage between the constant light (C(L)) and variable heavy domains has been reported previously. Transfectomas developed by introducing the expression construct of the amplifiable gene in dihydrofolate reductase- deficient Chinese hamster ovary (CHO dhfr-) cells secrete a 160 kDa homodimeric molecule, SCIgcCC49. The in vitro antigen binding properties of SCIgcCC49 are comparable to those of cCC49 and SCIgcCC49ΔC(H)1, a single- chain Ig deficient in constant heavy chain-1 (C(H)1) and C(L) domains. The antibody-dependent cellular cytotoxicity (ADCC) of SCIgcCC49 and cCC49 were also comparable. This single-gene approach for generating an immunoglobulin molecule may facilitate in vivo gene inoculation as well as ex vivo transfection of patients' cultured tumor-infiltrating lymphocytes for immunotherapy protocols for a variety of diseases, including cancer.
Keyword
Antitumor antibodyTAG-72 AntigenCarcinomaSingle geneSingle-chain IgEffector function
ISSN
0161-5890
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/S0161-5890(98)00109-6
Type
Article
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1. Journal Articles > Journal Articles
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