A human kringle domain-based fluorescence-linked immunosorbent assay system

Cited 5 time in webofscience Cited 3 time in scopus
  • Hit : 274
  • Download : 0
As a non-immunoglobulin protein scaffold, human kringle domain (KD) has attractive properties such as high specificity, stability, and production in bacterial hosts. Here, we developed a rapid and sensitive fluorescence-linked immunosorbent assay (FLISA) system using a fluorescent kringle domain (fluoKD), a fusion protein of a green fluorescent protein (GFP), and a kringle domain variant (KD548). Two kinds of fluoKDs in which MD was fused to the N terminus of GFP (N-fluoKD) or the C terminus of GFP (C-fluoKD) were constructed and characterized. In Escherichia coil host, both fluoKDs were produced in high yield and solubility and were successfully purified by a simple procedure. The purified fluoKDs exhibited strong fluorescent activities and high affinities to the target antigen. Furthermore, it was successfully demonstrated that the FL1SA with purified fluoKDs allowed for more rapid detection of target antigens with higher sensitivity compared with conventional enzyme-linked immunosorbent assay (ELISA), indicating that a simple, rapid, and sensitive immunoassay system could be developed by using MD instead of antibody or antibody fragments.
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
Issue Date
2014-04
Language
English
Article Type
Article
Keywords

PROTEIN SCAFFOLDS; ESCHERICHIA-COLI; BINDING-PROTEINS; ANTIBODIES; DISCOVERY

Citation

ANALYTICAL BIOCHEMISTRY, v.451, pp.63 - 68

ISSN
0003-2697
DOI
10.1016/j.ab.2014.01.019
URI
http://hdl.handle.net/10203/193054
Appears in Collection
CBE-Journal Papers(저널논문)
Files in This Item
There are no files associated with this item.
This item is cited by other documents in WoS
⊙ Detail Information in WoSⓡ Click to see webofscience_button
⊙ Cited 5 items in WoS Click to see citing articles in records_button

qr_code

  • mendeley

    citeulike


rss_1.0 rss_2.0 atom_1.0