Expression of foreign genes in mammalian cells using an antibody fusion system.
An expression system is described whereby a gene product is expressed fused to an antibody Fab fragment to form an antibody-like molecule. The antigen binding function of the original antibody is retained and the foreign gene replaces the CH2 and CH3 regions of the heavy chain. The fusion protein is secreted as if it were an antibody, and can be purified using the antigen-binding function of the Fab-like part of the molecule. In principle, any open reading frame can be expressed and it is not necessary to develop an individual purification scheme, or any analytical reagents such as antibodies, for the expressed protein, as both these functions can be performed by the Fab part of the fusion protein. In practice, the nature of the nonantibody part of the fusion influences the efficiency of expression and secretion, and detailed guidance is given on trouble-shooting and maximizing expression.
Forster
S J
SJ
Scotgen Biopharmaceuticals Inc., Aberdeen, UK.
Carr
F J
FJ
Harris
W J
WJ
Hamilton
A A
AA
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D001709:Biotechnology; D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D002846:Chromatography, Affinity; D003001:Cloning, Molecular / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D007140:Immunoglobulin Fab Fragments / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification; D014162:Transfection
D000906:Antibodies; D007140:Immunoglobulin Fab Fragments; D011993:Recombinant Fusion Proteins