Mehdi Golchin, Fatemeh Noori, Ali Akbar Khalili-Yazdi,
Volume 67, Issue 12 (3-2010)
Abstract
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Background: Recombinant antibodies are new versions of
monoclonal antibodies that are produced by recent molecular biology techniques.
These antibodies can be isolated by phage
display technology from immune or non-immune libraries. Recombinant
antibodies are applied to treatment of some diseases and also are increasingly
used for diagnosis and detection of many antigens. In
the latter case, the presence of antigen-antibody complexes has to be detected
by further approaches. The aim of current
research was to stain an anti-K99 phage antibody with
two different protein dyes and to apply them directly for detection of E.
coli K99
fimbriae.
Methods: In order to stain above antibody, a phagmid
vector carrying the anti-K99 single-chain Fv
(scFv) antibody was isolated, purified and
transformed into TG1 strain of E.
coli. Afterward,
the antibody was expressed in this cell as phage-scFv
antibody. Phage antibodies were
subsequently eluted, purified and stained with Disperse
Red dye 60
and Coomassie Brilliant
Blue. Finally,
the binding activity of coloured phage antibodies towards the purified K99
fimbriae was verified by immunoblotting.
Results: The results showed that anti-K99
phage antibody was stained with both dyes and the coloured phages were able to
recognize the corresponding antigen.
Conclusions: These protein stains that they usually do not
alter the protein structure can be used for staining phage antibodies. The
coloured phage antibodies retain their binding affinity for the antigens, and
therefore can be applied to detection of relevant antigens.