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Improving scFv antibody expression levels in the plant cytosol 1
Author(s) -
Schouten Alexander,
Roosien Jan,
de Boer Jan M,
Wilmink Annemiek,
Rosso Marie-Noëlle,
Bosch Dirk,
Stiekema Willem J,
Gommers Fred J,
Bakker Jaap,
Schots Arjen
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)01129-0
Subject(s) - kdel , cytosol , chemistry , microbiology and biotechnology , protoplast , subcellular localization , antibody , biochemistry , biology , enzyme , cytoplasm , gene , recombinant dna , genetics
Expression of single‐chain antibody fragments (scFvs) in the plant cytosol is often cumbersome. It was unexpectedly shown that addition at the C‐terminus of the ER retention signal KDEL resulted in significantly improved expression levels. In this report the cytosolic location of the scFv‐CK was confirmed, excluding possible mistranslocation to other subcellular compartments. It was shown that expression of several other scFvs was also improved in tobacco protoplasts. In addition expression was improved in transgenic potato. Changing from KDEL to KDEI did not affect the enhanced protein expression level. Addition of the KDEL motif is a simple and straightforward tool to stabilize in planta cytosolic expression of many scFvs.