Autor: Esperanza Torres, Carmen Vaquero, Liz Nicholson, Markus Sack, Eva Stöger, Jürgen Drossard, Paul Christou, Rainer Fischer, Yolande Perrin
Rok vydání: 1999
Předmět:
Zdroj: Transgenic Research. 8:441-449
ISSN: 0962-8819
DOI: 10.1023/a:1008969031219
Popis: We investigated the suitability of transformed rice cell lines as a system for the production of therapeutic recombinant antibodies. Expression constructs encoding a single-chain Fv fragment (scFvT84.66, specific for CEA, the carcinoembryonic antigen present on many human tumours) were introduced into rice tissue by particle bombardment. We compared antibody production levels when antibodies were either secreted to the apoplast or retained in the endoplasmic reticulum (ER) using a KDEL retention signal. Production levels were up to 14 times higher when antibodies were retained in the ER. Additionally, we compared constructs encoding different leader peptides (plant codon optimised murine immunoglobulin heavy and light chain leader peptides) and carrying alternative 5′ untranslated regions (the petunia chalcone synthase gene 5′ UTR and the tobacco mosaic virus omega sequence). We observed no significant differences in antibody production levels among cell lines transformed with these constructs. The highest level of antibody production we measured was 3.8 μg g−1 callus (fresh weight). Immunological analysis of transgenic rice callus confirmed the presence of functional scFvT84.66. We discuss the potential merits of cell culture for the production of recombinant antibodies and other valuable macromolecules.
Databáze: OpenAIRE
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