Improving scFv antibody expression levels in the plant cytosol

A. Schouten; J. Roosien; J.M. de Boer; A. Wilmink; M.N. rosso; D. Bosch; W.J. Stiekema; F.J. gommers; J. Bakker; A. Schots

doi:10.1016/S0014-5793(97)01129-0

Improving scFv antibody expression levels in the plant cytosol

A. Schouten, J. Roosien, J.M. de Boer, A. Wilmink, M.N. rosso, D. Bosch, W.J. Stiekema, F.J. gommers, J. Bakker, A. Schots

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71 Citations (Scopus)

Abstract

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 plants cytosolic expression of many scFvs.

Original language	English
Pages (from-to)	235-241
Journal	FEBS Letters
Volume	415
Issue number	2
DOIs	https://doi.org/10.1016/S0014-5793(97)01129-0
Publication status	Published - 1997

Keywords

KDEL retention signal
Localization
N-linked glycosylation
Protein expression
Single-chain antibody fragment
Transgenic plant

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title = "Improving scFv antibody expression levels in the plant cytosol",

abstract = "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 plants cytosolic expression of many scFvs.",

keywords = "KDEL retention signal, Localization, N-linked glycosylation, Protein expression, Single-chain antibody fragment, Transgenic plant",

author = "A. Schouten and J. Roosien and {de Boer}, J.M. and A. Wilmink and M.N. rosso and D. Bosch and W.J. Stiekema and F.J. gommers and J. Bakker and A. Schots",

year = "1997",

doi = "10.1016/S0014-5793(97)01129-0",

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journal = "FEBS Letters",

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T1 - Improving scFv antibody expression levels in the plant cytosol

AU - Schouten, A.

AU - Roosien, J.

AU - de Boer, J.M.

AU - Wilmink, A.

AU - rosso, M.N.

AU - Bosch, D.

AU - Stiekema, W.J.

AU - gommers, F.J.

AU - Bakker, J.

AU - Schots, A.

PY - 1997

Y1 - 1997

N2 - 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 plants cytosolic expression of many scFvs.

AB - 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 plants cytosolic expression of many scFvs.

KW - KDEL retention signal

KW - Localization

KW - N-linked glycosylation

KW - Protein expression

KW - Single-chain antibody fragment

KW - Transgenic plant

U2 - 10.1016/S0014-5793(97)01129-0

DO - 10.1016/S0014-5793(97)01129-0

M3 - Article

SN - 0014-5793

VL - 415

SP - 235

EP - 241

JO - FEBS Letters

JF - FEBS Letters

IS - 2

ER -

Improving scFv antibody expression levels in the plant cytosol

Abstract

Keywords

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