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Random mutagenesis of Bo NT /E H c nanobody to construct a secondary phage‐display library
Author(s) -
Shahi B.,
Mousavi Gargari S.L.,
Rasooli I.,
Rajabi Bazl M.,
Hoseinpoor R.
Publication year - 2014
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12526
Subject(s) - library science , medicine , computer science
Abstract 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 (Bo NT /E) was amplified by error‐prone PCR . Several biopanning rounds were used to screen the phage‐displaying Bo NT /E Hc nanobodies. The final nanobody, SHMR 4, with increased affinity recognized Bo NT /E toxin with no cross‐reactivity with other antigens especially with related Bo NT 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.