Production of antibody derivatives in the methylotrophic yeast Pichia pastoris.
New antibody derivatives are continuously being generated to interact with a range of therapeutic targets. The cost-effective and efficient production of these and other antibody derivatives is crucial for their further success. Here, we describe the construction of the expression vectors needed for heterologous expression of a Fab fragment in the yeast Pichia pastoris. The experimental conditions for lab-scale expressions are discussed, and an overview of an efficient purification strategy is presented.
Department of Cellular and Molecular Immunology, Vrije Universiteit Brussel (CMIM-VUB), Brussels, Belgium. sschoono@vub.ac.be
Steve
S
Schoonooghe
Jannick
J
Leoen
Jurgen
J
Haustraete
D000906:Antibodies / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors; D006639:Histidine / Q000302:isolation & purification; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis* / Q000302:isolation & purification; D000432:Methanol / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D009842:Oligopeptides / Q000302:isolation & purification; D010843:Pichia / Q000378:metabolism*
D000906:Antibodies; C471213:His-His-His-His-His-His; D007140:Immunoglobulin Fab Fragments; D009842:Oligopeptides; D006639:Histidine; D000432:Methanol