Functional improvement of antibody fragments using a novel phage coat protein III fusion system.
Functional expressions of proteins often depend on the presence of host specific factors. Frequently recombinant expression strategies of proteins in foreign hosts, such as bacteria, have been associated with poor yields or significant loss of functionality. Improvements in the performance of heterologous expression systems will benefit present-day quests in structural and functional genomics where high amounts of active protein are required. One example, which has been the subject of considerable interest, is recombinant antibodies or fragments thereof as expressions of these in bacteria constitute an easy and inexpensive method compared to hybridoma cultures. Such approaches have, however, often suffered from low yields and poor functionality. A general method is described here which enables expressions of functional antibody fragments when fused to the amino-terminal domain(s) of the filamentous phage coat protein III. Furthermore, it will be shown that the observed effect is neither due to improved stability nor increased avidity.
Jensen
Kim Bak
KB
Department of Molecular Biology, University of Aarhus, Aarhus, Denmark.
Larsen
Martin
M
Pedersen
Jesper Søndergaard
JS
Christensen
Peter Astrup
PA
Alvarez-Vallina
Luis
L
Goletz
Steffen
S
Clark
Brian F C
BF
Kristensen
Peter
P
D001483:Base Sequence; D036022:Capsid Proteins / Q000737:chemistry / Q000276:immunology*; D002850:Chromatography, Gel; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D007128:Immunoglobulin Fragments / Q000276:immunology*; D017912:Inovirus / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification
D036022:Capsid Proteins; D017931:DNA Primers; D007128:Immunoglobulin Fragments; D011993:Recombinant Fusion Proteins