title: Rice cell culture as an alternative production system for functional diagnostic and therapeutic antibodies.
abstract: 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 construct sencoding different leader peptides (plant codon optimised murine immunoglobulin heavy and light chain leader peptides from mAb24) 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 micrograms 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.
authors.affiliation: Molecular Biotechnology Unit, John Innes Centre, Norwich, UK.
authors.forename: E
authors.initials: E
authors.lastname: Torres
authors.forename: C
authors.initials: C
authors.lastname: Vaquero
authors.forename: L
authors.initials: L
authors.lastname: Nicholson
authors.forename: M
authors.initials: M
authors.lastname: Sack
authors.forename: E
authors.initials: E
authors.lastname: Stöger
authors.forename: J
authors.initials: J
authors.lastname: Drossard
authors.forename: P
authors.initials: P
authors.lastname: Christou
authors.forename: R
authors.initials: R
authors.lastname: Fischer
authors.forename: Y
authors.initials: Y
authors.lastname: Perrin
mesh_terms: D020121:5' Untranslated Regions; D000217:Acyltransferases / Q000235:genetics; D020816:Amino Acid Motifs; D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000378:metabolism; D002272:Carcinoembryonic Antigen / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics; D051379:Mice; D012275:Oryza / Q000166:cytology / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014027:Tobacco Mosaic Virus / Q000235:genetics
chemical_list: D020121:5' Untranslated Regions; D000906:Antibodies; D002272:Carcinoembryonic Antigen; D007135:Immunoglobulin Variable Region; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D000217:Acyltransferases; C020141:flavanone synthetase