An active single-chain antibody containing a cellulase linker domain is secreted by Escherichia coli.
Single-chain antibodies consist of the variable, antigen-binding domains of antibodies joined to a continuous polypeptide by genetically engineered peptide linkers. We have used the flexible interdomain linker region of a fungal cellulase to link together the variable domains of an anti-2-phenyloxazolone IgG1 and show here that the resulting single-chain antibody is efficiently secreted and released to the culture medium of Escherichia coli. The yield of affinity-purified single-chain antibody is 1-2 mg/l of culture medium and its affinity and stability are comparable to those of the corresponding native IgG.
Takkinen
K
K
VTT Biotechnical Laboratory, Espoo, Finland.
Laukkanen
M L
ML
Sizmann
D
D
Alfthan
K
K
Immonen
T
T
Vanne
L
L
Kaartinen
M
M
Knowles
J K
JK
Teeri
T T
TT
D000595:Amino Acid Sequence; D000918:Antibody Specificity; D001483:Base Sequence; D043366:Cellulose 1,4-beta-Cellobiosidase; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics*; D006026:Glycoside Hydrolases / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D010081:Oxazolone / Q000031:analogs & derivatives / Q000276:immunology; D011487:Protein Conformation; D011489:Protein Denaturation; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D014242:Trichoderma / Q000201:enzymology / Q000235:genetics*
D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; C018361:2-phenyloxazolone; D010081:Oxazolone; D006026:Glycoside Hydrolases; D043366:Cellulose 1,4-beta-Cellobiosidase