کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1183162 1492083 2016 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Affinity improvement by fine tuning of single-chain variable fragment against aflatoxin B1
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Affinity improvement by fine tuning of single-chain variable fragment against aflatoxin B1
چکیده انگلیسی


• A deleterious mutation was identified in an affinity-maturated scFv against aflatoxin B1.
• The back mutation of the deleterious residue resulted in a 3.2-fold affinity improvement.
• Specificity and structural properties were not influenced by the back mutation.

Aflatoxin B1 (AFB1) produced in Aspergillus flavus is a major hepatocarcinogen found in foods and feed. For effective immunological detection of AFB1 at low concentrations, the development of high affinity antibody for AFB1 is required. Previously, an affinity-maturated single-chain variable fragment containing 6 mutations (scFv-M37) was isolated from an artificial mutagenic library, which showed a 9-fold higher affinity than its wild type scFv. In this study, the effect of the 6 mutated residues on the affinity improvement was characterized using surface plasmon resonance analysis, which identified a deleterious mutation (VH-A110T) located on a framework region of the scFv-M37. The back mutation of VH-A110T resulted in a 3.2-fold affinity improvement, which was attributed to decrease of dissociation rate constant (kd) in interaction between AFB1 and the back mutant scFv. The biophysical analyses using circular dichroism and gel filtration revealed that the back mutation of VH-A110T caused a subtle conformational change of the scFv toward tighter binding to AFB1.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Food Chemistry - Volume 209, 15 October 2016, Pages 312–317
نویسندگان
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