Affinity maturation of single-chain variable fragment specific for aflatoxin B(1) using yeast surface display.
Food Chem
; 188: 604-11, 2015 Dec 01.
Article
in En
| MEDLINE
| ID: mdl-26041237
As aflatoxin B1 is one of the most toxic mycotoxins, it is important to detect and to quantify aflatoxin B1 accurately by immunological methods. To enhance aflatoxin B1-binding affinity of the single-chain variable fragment, yeast surface display technique combined with fluorescence-activated cell sorting was applied. A randomly mutated scFv library was subjected to 4 rounds of fluorescence-activated cell sorting, resulting in isolation of 5 scFv variants showing an affinity improvement compared to the parental wild type scFv. The best scFv with a 9-fold improvement in affinity for aflatoxin B1 exhibited similar specificity to the monoclonal antibody. Most of the mutations in scFv-M37 were located outside of the canonical antigen-contact loops, suggesting that its affinity improvement might be driven by an allosteric effect inducing scFv-M37 to form a more favorable binding pocket for aflatoxin B1 than the wild type scFv.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Aspergillus
/
Aflatoxin B1
/
Single-Chain Antibodies
/
Flow Cytometry
Language:
En
Journal:
Food Chem
Year:
2015
Document type:
Article
Country of publication:
Reino Unido