Improving scFv antibody expression levels in the plant cytosol

FEBS Letters

Gommers, Fred J²  Roosien, Jan²  Schots, Arjen²  Rosso, Marie-Noëlle¹  Schouten, Alexander²  Stiekema, Willem J³  Wilmink, Annemiek³  Bosch, Dirk³  de Boer, Jan M²  Bakker, Jaap² 

[1] Laboratoire de Biologie des Invertébrés, INRA, P.O. Box 2078, 06606 Antibes, France;Department of Nematology, Wageningen Agricultural University, P.O. Box 8123, 6700 ES Wageningen, The Netherlands;Department of Molecular Biology, Centre for Plant Breeding and Reproduction Research (CPRO-DLO), P.O. Box 16, 6700 AA Wageningen, The Netherlands

关键词: Localization;    KDEL retention signal;    N-linked glycosylation;    Protein expression;    Single-chain antibody fragment;    Transgenic plant;    EDTA;    ethylenediaminetetraacetic acid;    ER;    endoplasmic reticulum;    HEPES;    N-(2-hydroxyethyl)piperazine-N′-(2-ethanesulfonic acid);    NP-40;    nonidet P-40;    PAGE;    polyacrylamide gel electrophoresis;    PCR;    polymerase chain reaction;    PVDF;    polyvinylidene difluoride;    rER;    rough endoplasmic reticulum;    SDS;    sodium dodecyl sulfate;   

DOI  :  10.1016/S0014-5793(97)01129-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown