New ELISA approach based on coiled-coil interactions

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Journal titleJournal of Immunological Methods
Pages161167; # of pages: 7
SubjectEGF; E/K coiled-coil interactions; ELISA test; Acids; analysis; Animal; antibody affinity; antibody detection; antibody specificity; article; Binding Sites; bio; Biotechnology; Canada; Cells; controlled study; Dimerization; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Escherichia coli; Gene Expression; genome; Humans; I; In Vitro; molecular interaction; monitoring; NPArC; Nucleic Acids; peptide analysis; Peptides; Phosphorylation; polyhistidine tag; priority journal; Protein; protein analysis; protein antibody; Protein Folding; protein protein interaction; Protein Structure; Secondary; Proteins; Purification; Recombinant protein; sensitivity analysis; Sensitivity and Specificity; synthesis; Tags; Tissue Engineering; Vitro
AbstractThe de novo designed heterodimeric E/K coiled-coil system has been previously demonstrated to be an excellent capture/dimerization system applicable to various needs in both biotechnology and pharmaceutical fields. Those include controlled protein dimerization, capture, purification and Western-blot detection. We here report the development of a new generation of ELISA test based on coiled-coil interactions for the direct quantitation of coil-tagged epidermal growth factor (EGF). The new approach was evaluated for its specificity, plate storability and reusability as well as for convenience when compared to commercially available systems. Our results show a similar affinity/sensitivity to standard capturing antibody-based ELISA systems and an improved affinity/sensitivity when compared to the commercially available Ni-NTA capture system. The E/K coiled-coil ELISA system was validated with respect to recovery, intra- and inter-assay variations. The practical working range was estimated to be between 5.2 and 34,000 pM. Furthermore, the storability and reusability of the plates was greater than the two aforementioned systems, suggesting that the E/K coiled-coil system is a good alternative to traditional tags such as poly-histidine for the development of ELISA tests aiming at quantitating coil-tagged proteins. -� 2010 Elsevier B.V
Publication date
AffiliationNRC Biotechnology Research Institute; National Research Council Canada
Peer reviewedYes
NRC number52766
NPARC number16616678
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Record identifierca2e232f-148e-47d1-a241-0c678651ab40
Record created2011-03-31
Record modified2016-05-09
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