| Download | - View accepted manuscript: New ELISA approach based on coiled-coil interactions (PDF, 739 KiB)
|
|---|
| DOI | Resolve DOI: https://doi.org/10.1016/j.jim.2010.09.027 |
|---|
| Author | Search for: Liberelle, Benoît; Search for: Bartholin, Laurence; Search for: Boucher, Cyril; Search for: Murschel, Frédéric; Search for: Jolicoeur, Mario; Search for: Durocher, Yves1; Search for: Merzouki, Abderrazzak; Search for: De Crescenzo, Gregory |
|---|
| Affiliation | - National Research Council of Canada. NRC Biotechnology Research Institute
|
|---|
| Format | Text, Article |
|---|
| Subject | EGF; E/K coiled-coil interactions; ELISA test; Acids; antibody affinity; antibody detection; antibody specificity; Binding Sites; biotechnology; Biotechnology; Canada; Cells; Dimerization; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Escherichia coli; Gene Expression; genome; In Vitro; molecular interaction; monitoring; NPArC; Nucleic Acids; peptide analysis; Peptides; Phosphorylation; polyhistidine tag; 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 |
|---|
| Abstract | The 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 | 2010-10-01 |
|---|
| In | |
|---|
| Language | English |
|---|
| Peer reviewed | Yes |
|---|
| NRC number | NRCC 52766 |
|---|
| NPARC number | 16616678 |
|---|
| Export citation | Export as RIS |
|---|
| Report a correction | Report a correction (opens in a new tab) |
|---|
| Record identifier | ca2e232f-148e-47d1-a241-0c678651ab40 |
|---|
| Record created | 2011-03-31 |
|---|
| Record modified | 2022-11-18 |
|---|
