[Construction of immune library of murine immunoglobulin genes and screening of single-chain Fv-antibodies specific to diphtheria toxin B subunit].
The aim of this work was to obtain the panel of recombinant single-chain Fv-antibodies against diphtheria toxin B subunit, the main diagnostic and pathogenic antigen of Corynebacterium diphtheriae. For this purpose we have constructed the immune library of murine immunoglobulin genes. A number of scFv specific to diphtheria toxin B subunit were acquired from the obtained library after one round of selection by phage-display. ScFv encoding DNA-fragments of eight clones were subcloned into plasmid pET-22b(+). It was shown that selected scFv were highly specific to diphtheria toxin B subunit, with affinity constant for different clones ranged from 10(7) to 10(9) M(-1).
Oliĭnyk
O S
OS
Kaberniuk
A A
AA
Redchuk
T A
TA
Korotkevych
N V
NV
Labyntsev
A Iu
AIu
Romaniuk
S I
SI
Kolybo
D V
DV
Komisarenko
S V
SV
D000818:Animals; D000915:Antibody Affinity; D000918:Antibody Specificity; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000276:immunology; D004165:Diphtheria / Q000175:diagnosis / Q000276:immunology; D004167:Diphtheria Toxin / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D019151:Peptide Library; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D057127:Single-Chain Antibodies / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D013154:Spleen / Q000166:cytology / Q000276:immunology
D018076:DNA, Complementary; D004167:Diphtheria Toxin; D019151:Peptide Library; D021122:Protein Subunits; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies