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Updated: Apr 30, 2026
Construction of Synthetic Phage Displayed Fab Library with Tailored Diversity
Published on: May 1, 2018
Random mutagenesis of BoNT/E Hc nanobody to construct a secondary phage-display library
B Shahi1, S L Mousavi Gargari, I Rasooli
1Department of Biology, Basic Science Faculty, Shahed University, Tehran, Iran.
Aims:
To construct secondary mutant phage-display library of recombinant single variable domain (VHH) against botulinum neurotoxin E by error-prone PCR.
Methods And Results:
The gene coding for specific VHH derived from the camel immunized with binding domain of botulinum neurotoxin E (BoNT/E) was amplified by error-prone PCR. Several biopanning rounds were used to screen the phage-displaying BoNT/E Hc nanobodies. The final nanobody, SHMR4, with increased affinity recognized BoNT/E toxin with no cross-reactivity with other antigens especially with related BoNT toxins.
Conclusions:
The constructed nanobody could be a suitable candidate for VHH-based biosensor production to detect the Clostridium botulinum type E.
Significance And Impact Of The Study:
Diagnosis and treatment of botulinum neurotoxins are important. Generation of high-affinity antibodies based on the construction of secondary libraries using affinity maturation step leads to the development of reagents for precise diagnosis and therapy.
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