High-level secretion of two antibody single chain Fv fragments by Pichia pastoris.
The diagnostic and therapeutic applications of antibody single-chain Fv (sFv) fragments often require large amounts of protein that can be problematic and expensive to obtain. Here we report the secretion of two sFv fragments by the yeast Pichia pastoris at levels up to 250 mg/l. Soluble sFv fragments were purified from culture supernatants in one step by affinity or metal-chelating chromatography, and were indistinguishable from their bacterially expressed counterparts in terms of affinity. Secretion of functional sFv fragments by Pichia pastoris provides a low cost, high yield alternative to current sFv expression systems.
Eldin
P
P
Department of Laboratory Medicine and Pathology, School of Medicine, University of Minnesota, Minneapolis 55455, USA.
Pauza
M E
ME
Hieda
Y
Y
Lin
G
G
Murtaugh
M P
MP
Pentel
P R
PR
Pennell
C A
CA
D000818:Animals; D000915:Antibody Affinity; D000918:Antibody Specificity; D019011:Antigens, CD7 / Q000276:immunology; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D005803:Genes, Immunoglobulin; D006825:Hybridomas; D007128:Immunoglobulin Fragments / Q000235:genetics*; D007135:Immunoglobulin Variable Region; D051379:Mice; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D011994:Recombinant Proteins
D019011:Antigens, CD7; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; C055966:immunoglobulin Fv