[Construction of combinatorial immune library of single chain human antibodies to orthopoxviruses and selection from this library antibodies to recombinant protein prA30L of variola virus].
A combinatorial immune library of human single-chain antibody fragments (scFv) was constructed on the base of genes encoding variable domains of heavy and light chains of immunoglobulins cloned from the lymphocytes of four vaccinia virus (VACV) vaccinated donors. The size of the library was 3 x 10(7) independent clones. After the library was enriched with the clones producing scFv against recombinant analogue of variola virus surface protein prA30L, a panel of unique antibodies specific to both prA30L and VACV was selected from the library. A plaque reduction neutralization test was performed for all selected antibodies and two antibodies were shown to be able to neutralize plaque formation of VACV in Vero E6 cells monolayer. Binding specificities of these antibodies were confirmed using ELISA and Western blot analysis. To determine the amino acid sequences of neutralizing antibodies their genes were sequenced.
Dubrovskaia
V V
VV
Ulitin
A B
AB
Laman
A G
AG
Gileva
I P
IP
Bormotov
N I
NI
Il'ichev
A A
AA
Brovko
F A
FA
Shchelkunov
S N
SN
Belanov
E F
EF
Tikunova
N V
NV
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology; D000914:Antibodies, Viral / Q000235:genetics* / Q000276:immunology; D000918:Antibody Specificity / Q000235:genetics / Q000276:immunology; D002522:Chlorocebus aethiops; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D012901:Variola virus / Q000235:genetics / Q000276:immunology*; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*; D038301:Virus Inactivation / Q000187:drug effects
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D014764:Viral Proteins