Format

Send to

Choose Destination
J Immunol Methods. 1999 Nov 19;230(1-2):121-30.

Fluobodies: green fluorescent single-chain Fv fusion proteins.

Author information

1
Laboratory of Monoclonal Antibodies, Wageningen University and Research Centre, P.O. Box 8123, 6700 ES, Wageningen, The Netherlands.

Abstract

An expression system (pSKGFP), which permits the expression of single-chain variable fragments as fusion proteins with modified green fluorescent proteins, was designed. This expression system is comparable to frequently used phage display vectors and allows single-step characterization of the selected recombinant antibodies by flow cytometry or fluorescent cell staining. Two different single-chain variable fragment antibodies, both directed against the lipopolysaccharide of the bacterium Ralstonia solanacearum have been genetically fused to a red-shifted green fluorescent protein and the produced fusion protein tested for usefulness. These fluobodies can be produced in cultures of bacterial cells and purified using immobilized metal affinity chromatography. They function well in flow cytometry and immunofluorescent cell staining, are specific for their target antigens and, unlike FITC-conjugated antibodies, they do not fade upon illumination.

PMID:
10594359
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Elsevier Science
Loading ...
Support Center