Affinity maturation of single-chain variable fragment specific for aflatoxin B-1 using yeast surface display

作者：Min Won Ki; Kim Sung Gun; Seo Jin Ho^*

来源：Food Chemistry, 2015, 188: 604-611.

DOI：10.1016/j.foodchem.2015.04.117

• Single-chain variable fragment
• Aflatoxin B-1
• Yeast surface display
• Fluorescence-activated cell sorting

As aflatoxin B-1 is one of the most toxic mycotoxins, it is important to detect and to quantify aflatoxin B-1 accurately by immunological methods. To enhance aflatoxin B-1-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 B-1 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 B-1 than the wild type scFv.

• 出版日期2015-12-1