Antibody production by the gram-positive bacterium Bacillus megaterium.
The increasing demand for recombinant antibodies as detection reagents in research, diagnostics, and therapy requires appropriate production systems. In contrast to antibody therapies, small recombinant antibody fragments like Fab and scFv are sufficient for most applications in research and diagnostics. These antibody fragments can also be produced in bacterial hosts. Gram-negative bacteria, particularly Escherichia coli, were extensively studied for the recombinant antibody production but they showed only a limited capacity to secrete antibody fragments into the medium--a prerequisite for easy downstream processing. Gram-positive bacteria are known to efficiently secrete recombinant proteins into the medium. Recently, we demonstrated the production of scFv and scFab fragments in Bacillus megaterium. Here, we describe the process in detail from transformation of B. megaterium to production and purification of scFv fragments.
Jordan
Eva
E
Institute of Biochemistry and Biotechnology, Technical University Braunschweig, Braunschweig, Germany.
Al-Halabi
Laila
L
Schirrmann
Thomas
T
Hust
Michael
M
D000906:Antibodies / Q000276:immunology*; D001410:Bacillus megaterium / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D007128:Immunoglobulin Fragments / Q000096:biosynthesis; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000302:isolation & purification; D018189:Immunomagnetic Separation; D008967:Molecular Biology / Q000379:methods*; D010957:Plasmids / Q000235:genetics; D011523:Protoplasts / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification; D014170:Transformation, Genetic
D000906:Antibodies; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D004247:DNA