Random Phage Display-Based Screening of Peptides that Bind to Botulinum Neurotoxin Binding Protein, Nontoxic Nonhemagglutinin

Sagane, Yoshimasa; Miyata, Keita; Kurihara, Sayuri; Yoneyama, T.; Inui, Ken Ichi; Miyashita, Shin-Ichiro; Hayashi, Shigenari; Suzuki, Tomonori; Niwa, Koichi

doi:10.1007/s00284-013-0355-0

Cited by 3 publications

(1 citation statement)

References 11 publications

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“…Specific antibodies play important roles in the diagnosis, treatment, and prevention of bacterial toxin virulence. Attempts have been made to develop artificial peptides for targeting bacterial toxins [19][20][21][22][23][24][25]. Vacuolating cytotoxin A (VacA) is produced by Helicobacter pylori, infection with which frequently causes chronic gastritis, peptic ulcers, and stomach cancer [26].…”

Section: Introductionmentioning

confidence: 99%

Complementary DNA display selection of high-affinity peptides binding the vacuolating toxin (VacA) ofHelicobacter pylori

et al. 2015

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Artificial peptides designed for molecular recognition of a bacterial toxin have been developed. Vacuolating cytotoxin A protein (VacA) is a major virulence factor of Helicobacter pylori, a gram-negative microaerophilic bacterium inhabiting the upper gastrointestinal tract, particularly the stomach. This study attempted to identify specific peptide sequences with high affinity for VacA using systematic directed evolution in vitro, a cDNA display method. A surface plasmon resonance-based biosensor and fluorescence correlation spectroscopy to examine binding of peptides with VacA identified a peptide (GRVNQRL) with high affinity. Cyclization of the peptide by attaching cysteine residues to both termini improved its binding affinity to VacA, with a dissociation constant (Kd ) of 58 nm. This study describes a new strategy for the development of artificial functional peptides, which are promising materials in biochemical analyses and medical applications.

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Section: Introductionmentioning

confidence: 99%

Complementary DNA display selection of high-affinity peptides binding the vacuolating toxin (VacA) ofHelicobacter pylori

et al. 2015

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Construction of “Toxin Complex” in a Mutant Serotype C Strain of Clostridium botulinum Harboring a Defective Neurotoxin Gene

et al. 2016

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A non-toxigenic mutant of the toxigenic serotype C Clostridium botulinum strain Stockholm (C-St), C-N71, does not produce the botulinum neurotoxin (BoNT). However, the original strain C-St produces botulinum toxin complex, in which BoNT is associated with non-toxic non-hemagglutinin (NTNHA) and three hemagglutinin proteins (HA-70, HA-33, and HA-17). Therefore, in this study, we aimed to elucidate the effects of bont gene knockout on the formation of the "toxin complex." Nucleotide sequence analysis revealed that a premature stop codon was introduced in the bont gene, whereas other genes were not affected by this mutation. Moreover, we successfully purified the "toxin complex" produced by C-N71. The "toxin complex" was identified as a mixture of NTNHA/HA-70/HA-17/HA-33 complexes with intact NTNHA or C-terminally truncated NTNHA, without BoNT. These results indicated that knockout of the bont gene does not affect the formation of the "toxin complex." Since the botulinum toxin complex has been shown to play an important role in oral toxin transport in the human and animal body, a non-neurotoxic "toxin complex" of C-N71 may be valuable for the development of an oral drug delivery system.

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Production of Recombinant Human scFv Against Tetanus Toxin Heavy Chain by Phage Display Technology

Khalili

Lakzaei

Rasaee

et al. 2015

Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

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Tetanus, as a major cause of death in developing countries, is caused by tetanus neurotoxin. Recombinant antibodies against tetanus neurotoxin can be useful in tetanus management. Phage display of antibody fragments from immune human antibody libraries with single chain constructs combining the variable fragments (scFv) has been one of the most prominent technologies in antibody engineering. The aim of this study was the generation of a single chain fragment of variable region (scFv) library and selection of specific antibodies with high affinity against tetanus toxin. Immune human single chain fragment variable (HuscFv) antibody phagemid library was displayed on pIII of filamentous bacteriophage. Selection of scFv clones was performed against tetanus toxin antigens after three rounds of panning. The selected scFv clones were analyzed for inhibition of tetanus toxin binding to ganglioside GT1b. After the third round of panning, over 35 HuscFv phages specific for tetanus toxin were isolated from this library of which 15 clones were found to bind specifically to tetanus toxin. The selected HuscFv phages expressed as a soluble HuscFv peptide and some clones showed positive signals against tetanus toxin. We found that six HuscFv clones inhibit toxin binding to ganglioside GT1b. These selected antibodies can be used in the management of tetanus.

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Random Phage Display-Based Screening of Peptides that Bind to Botulinum Neurotoxin Binding Protein, Nontoxic Nonhemagglutinin

Cited by 3 publications

References 11 publications

Complementary DNA display selection of high-affinity peptides binding the vacuolating toxin (VacA) ofHelicobacter pylori

Complementary DNA display selection of high-affinity peptides binding the vacuolating toxin (VacA) ofHelicobacter pylori

Construction of “Toxin Complex” in a Mutant Serotype C Strain of Clostridium botulinum Harboring a Defective Neurotoxin Gene

Production of Recombinant Human scFv Against Tetanus Toxin Heavy Chain by Phage Display Technology

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