Back to Search
Start Over
Generation of functional single-chain fragment variable from hybridoma and development of chemiluminescence enzyme immunoassay for determination of total malachite green in tilapia fish.
- Source :
-
Food chemistry [Food Chem] 2021 Feb 01; Vol. 337, pp. 127780. Date of Electronic Publication: 2020 Aug 05. - Publication Year :
- 2021
-
Abstract
- To determine malachite green (MG) and its major metabolite, leucomalachite green (LMG) residual levels in tilapia fish, chemiluminescent enzyme immunoassay (CLEIA) was developed based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein. At first, V <subscript>H</subscript> and V <subscript>L</subscript> gene sequences were cloned from hybridoma cell lines secreting monoclonal antibody against LMG, and then thoroughly by database-assisted sequence analysis. Finally, the productive V <subscript>H</subscript> and V <subscript>L</subscript> were assembled to an intact scFv sequence and engineered to produce scFv-AP fusion protein. The fusion protein was further identified as a bifunctional reagent for immunoassay, then a sensitive one-step CLEIA against LMG was developed with a half-maximal inhibitory concentration (IC <subscript>50</subscript> ) and limit of detection (LOD) of 1.3 and 0.04 ng/mL, respectively. The validation results of this novel competitive CLEIA was in line with those obtained by classical HPLC method for determination of total MG in spiked and field incurred samples.<br />Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.<br /> (Copyright © 2020 Elsevier Ltd. All rights reserved.)
Details
- Language :
- English
- ISSN :
- 1873-7072
- Volume :
- 337
- Database :
- MEDLINE
- Journal :
- Food chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 32799164
- Full Text :
- https://doi.org/10.1016/j.foodchem.2020.127780