[Characterization of a panel of mouse single-chain antibodies against recombinant human interferon beta1b].
A panel of single-chain antibodies (ScFv's--single-chain Fv-antibodies) against recombinant human interferon beta 1b (rhIFN-beta1b) has been obtained from immune and naive combinatorial cDNA libraries of the mouse variable immunoglobulin genes. ScFv's were expressed into Escherichia coli cells. For producers isolated from the immune library a difference in production yield of ScFv's in periplasm and incubation medium as well as their expression stability in passages and storage stability have been demonstrated. After sequencing of target DNA the multiple alignment and structural analysis of ScFv's sequences with different primary structures were carried out and significant difference in both complementarity-determining (CDR) and framework (FR) regions of their variable domains has been shown. For the ScFv's isolated from the immune library, specificity of their binding with native and denatured rhIFN-beta1b in ELISA and Western-blotting as well as their high storage stability have been shown. The affinity constants for each representatives of the ScFv's panel were in the range from 1.96 x 10(-8) to 1.69 x 10(-9) M.
Pavlova
M V
MV
Nikolaev
Iu S
IuS
Irodov
D M
DM
Okunev
O V
OV
Kordium
V A
VA
Gil'chuk
P V
PV
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D006801:Humans; D015151:Immunoblotting; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D007147:Immunoglobulin Light Chains / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D000068576:Interferon beta-1b; D016899:Interferon-beta / Q000276:immunology*; D051379:Mice
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; C055966:immunoglobulin Fv; D000068576:Interferon beta-1b; D016899:Interferon-beta