Very high expression of an anti-carcinoembryonic antigen single chain Fv antibody fragment in the yeast Pichia pastoris.
In this paper we report the development of a recombinant strain of the yeast Pichia pastoris, which secretes an anti-carcinoembryonic antigen single chain Fv (scFv) antibody fragment to the culture supernatant as a biologically active protein, at levels of 1.2 g l(-1). The yeast scFv was purified by IMAC, with a final yield of approximately 0.440 g of 93% pure scFv per liter of culture supernatant. The specific activity in ELISA of the yeast scFv was almost three times higher than that of a bacterial periplasmic counterpart. These results reaffirm that the yeast P. pastoris is a suitable host for high level production of scFv antibody fragments with potential in vivo diagnostic and therapeutic applications.
Freyre
F M
FM
Division of Immunotechnology and Diagnostics, Center for Genetic Engineering and Biotechnology, La Habana, Cuba.
Vázquez
J E
JE
Ayala
M
M
Canaán-Haden
L
L
Bell
H
H
Rodríguez
I
I
González
A
A
Cintado
A
A
Gavilondo
J V
JV
D000251:Adenosine Triphosphatases; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001709:Biotechnology / Q000379:methods; D002272:Carcinoembryonic Antigen / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D004268:DNA-Binding Proteins; D004797:Enzyme-Linked Immunosorbent Assay; D029968:Escherichia coli Proteins; D005285:Fermentation; D015966:Gene Expression Regulation, Fungal; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D051379:Mice; D051719:MutS DNA Mismatch-Binding Protein; D010843:Pichia / Q000235:genetics / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D014170:Transformation, Genetic
D001426:Bacterial Proteins; D002272:Carcinoembryonic Antigen; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; D000251:Adenosine Triphosphatases; D051719:MutS DNA Mismatch-Binding Protein; C497175:MutS protein, E coli