[Preparation of single-chained antibodies to human ferritin in Escherichia coli].
We have cloned cDNA copies of the immunoglobulin heavy and light chain genes from hybridoma cells able to produce antibody against human ferritin. Variable segments of these genes were obtained using the polymerase chain reaction (PCR). The specific amplifications of ligase reaction products were carried out to combine the variable segments with DNA fragments coding for a peptide-linker and for a signal peptide of the cloned pelB gene of Erwinia carotovora. During the antiferritin single-chain antibody gene expression under the T7 RNA polymerase control in E. coli the processed molecules of recombinant proteins formed aggregates in periplasm. The reversible denaturation in the absence of reducing agents had allowed us to obtain single-chain antibodies with the original binding specificity toward human ferritin.
Bespalov
I A
IA
Shiianov
P A
PA
Lukashevich
L V
LV
Lunev
V E
VE
Tribush
S S
SS
Gaponova
G I
GI
Deev
S M
SM
D000595:Amino Acid Sequence; D000906:Antibodies / Q000235:genetics* / Q000276:immunology; D000918:Antibody Specificity; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA; D004926:Escherichia coli; D005293:Ferritins / Q000276:immunology*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D000906:Antibodies; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D004247:DNA; D005293:Ferritins
Poluchenie odnotsepochechnykh antitel k ferritinu cheloveka v Escherichia coli.