Type

Journal Article

Authors

Montarop Yamabhai
Florian Rüker
Richard O'Kennedy
Kiattawee Choowongkomon
Kuntalee Rangnoi

Subjects

Microbiology

Topics
chemistry ligands immunology peptide library genetics dna shuffling single chain antibodies humans aflatoxin b1 enzyme linked immunosorbent assay molecular docking simulation

Enhancement and Analysis of Human Antiaflatoxin B1 (AFB1) scFv Antibody-Ligand Interaction Using Chain Shuffling. (2018)

Abstract A human antiaflatoxin B1 (AFB1) scFv antibody (yAFB1-c3), selected from a naı̈ve human phage-displayed scFv library, was used as a template for improving and analysis of antibody-ligand interactions using the chain-shuffling technique. The variable-heavy and variable-light (VH/VL)-shuffled library was constructed from the VH of 25 preselected clones recombined with the VL of yAFB1-c3 and vice versa. Affinity selection from these libraries demonstrated that the VH domain played an important role in the binding of scFv to free AFB1. Therefore, in the next step, VH-shuffled scFv library was constructed from variable-heavy (VH) chain repertoires, amplified from the naı̈ve library, recombined with the variable-light (VL) chain of the clone yAFB1-c3. This library was then used to select a specific scFv antibody against soluble AFB1 by a standard biopanning method. Three clones that showed improved binding properties were isolated. Amino acid sequence analysis indicated that the improved clones have amino acid mutations in framework 1 (FR1) and the complementarity determining region (CDR1) of the VH chain. One clone, designated sAFH-3e3, showed 7.5-fold improvement in sensitivity over the original scFv clone and was selected for molecular binding studies with AFB1. Homology modeling and molecular docking were used to compare the binding of this and the original clones. The results confirmed that VH is more important than VL for AFB1 binding.
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Full list of authors on original publication

Montarop Yamabhai, Florian Rüker, Richard O'Kennedy, Kiattawee Choowongkomon, Kuntalee Rangnoi

Experts in our system

1
R O'Kennedy
Dublin City University
Total Publications: 211