Bacterial and plant‐produced scFv proteins have similar antigen‐binding properties

FEBS Letters

Depicker, Ann¹  De Neve, Myriam¹  De Wilde, Chris¹  De Jaeger, Geert¹  Van Montagu, Marc¹  Bruyns, Anne-Marie¹ 

[1] Laboratorium voor Genetica, via the Department of Genetics, affiliated to the Flanders Interuniversity Institute for Biotechnology, Universiteit Gent, K.L. Ledeganckstraat 35, B-9000 Gent, Belgium

关键词: Antibody affinity;    Escherichia coli;    Heterologous gene expression;    Nicotiana tabacum;    Single-chain variable fragment;    Transgenic plant;    EDTA;    ethylenediaminetetraacetic acid;    ER;    endoplasmic reticulum;    PCR;    polymerase chain reaction;    PMSF;    phenylmethylsulphonyl fluoride;    SDS;    sodium dodecyl sulphate;    TSP;    total soluble protein;   

DOI  :  10.1016/0014-5793(96)00372-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A gene encoding a single-chain variable (scFv) antibody fragment was expressed as a cytoplasmic and endoplasmic reticulum-targeted protein in transgenic tobacco plants. In both cases, the scFv accumulated up to 0.01% of total soluble protein (TSP). The same scFv fragment was also produced in the periplasm of Escherichia coli. Measurement of the affinity by ELISA indicates that the affinity of the bacterially made scFv is about 80-fold lower than that of the parental F_ab fragment. The results suggest that the affinity of the plant-produced scFv fragments is reduced to a similar extent, implying that all the plant-produced scFv fragments are antigen binding.

【 授权许可】

Unknown