Production of single chain fragment variable (scFv) antibodies in Escherichia coli using the LEX™ bioreactor.
For proteome research, antibodies against a growing number of antigens must be generated and characterized. The high throughput generation of antibody fragments, using in vitro selection, requires bacterial expression of antibody fragments. This created a need to establish an expression method to improve the parallel production of many antibody fragments. In this study, we describe the development of a high throughput bacterial production method for single chain fragment variables (scFvs) using shaking flasks or the LEX™ bioreactor. We compared the influence of a set of production parameters on Escherichia coli production of four different scFv. The results led to an optimized protocol for the parallel production of multiple antibody fragments.
Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie, Spielmannstr. 7, 38106 Braunschweig, Germany.
Sebastian
S
Miethe
Torsten
T
Meyer
Stefanie
S
Wöhl-Bruhn
André
A
Frenzel
Thomas
T
Schirrmann
Stefan
S
Dübel
Michael
M
Hust
D019149:Bioreactors / Q000382:microbiology*; D001709:Biotechnology / Q000295:instrumentation* / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
D011993:Recombinant Fusion Proteins; D057127:Single-Chain Antibodies