Production of Recombinant Human scFv Against Tetanus Toxin Heavy Chain by Phage Display Technology.
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.
1 Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences , Tehran, Iran .
Ehsan
E
Khalili
1 Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences , Tehran, Iran .
Mostafa
M
Lakzaei
2 Department of Medical Biotechnology, School of Medical Sciences, Tarbiat Modares University , Tehran, Iran .
Mohhamad Javad
MJ
Rasaee
1 Department of Clinical Biochemistry, School of Medicine, Tehran University of Medical Sciences , Tehran, Iran .|3 Recombinant Vaccine Research Center, Tehran University of Medical Sciences , Tehran, Iran .
Mahdi
M
Aminian
D000942:Antigens, Bacterial / Q000276:immunology*; D001435:Bacteriophages / Q000276:immunology*; D061505:Cell Surface Display Techniques / Q000379:methods; D005732:Gangliosides / Q000276:immunology; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000276:immunology*; D008666:Metalloendopeptidases / Q000276:immunology; D019151:Peptide Library; D011994:Recombinant Proteins / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000276:immunology*; D013742:Tetanus / Q000276:immunology*; D013744:Tetanus Toxin / Q000276:immunology*
D000942:Antigens, Bacterial; D005732:Gangliosides; D007143:Immunoglobulin Heavy Chains; D019151:Peptide Library; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D013744:Tetanus Toxin; C035979:tetanospasmin; C016481:trisialoganglioside GT1; D008666:Metalloendopeptidases