On the influence of vector design on antibody phage display.
Phage display technology is an established technology particularly useful for the generation of monoclonal antibodies (mAbs). The isolation of phagemid-encoded mAb fragments depends on several features of a phage preparation. The aims of this study were to optimize phage display vectors, and to ascertain if different virion features can be optimized independently of each other. Comparisons were made between phagemid virions assembled by g3p-deficient helper phage, Hyperphage, Ex-phage or Phaberge, or corresponding g3p-sufficient helper phage, M13K07. All g3p-deficient helper phage provided a similar level of antibody display, significantly higher than that of M13K07. Hyperphage packaged virions at least 100-fold more efficiently than did Ex-phage or Phaberge. Phaberge's packaging efficiency improved by using a SupE strain. Different phagemids were also compared. Removal of a 56 base pair fragment from the promoter region resulted in increased display level and increased virion production. This critical fragment encodes a lacZ'-like peptide and is also present in other commonly used phagemids. Increasing display level did not show statistical correlation with phage production, phage infectivity or bacterial growth rate. However, phage production was positively correlated to phage infectivity. In summary, this study demonstrates simultaneously optimization of multiple and independent features of importance for phage selection.
Soltes
Glenn
G
Cangene Corporation, 3403 American Drive, Mississauga, Ontario L4V 1T4, Canada.
Hust
Michael
M
Ng
Kitty K Y
KK
Bansal
Aasthaa
A
Field
Johnathan
J
Stewart
Donald I H
DI
Dübel
Stefan
S
Cha
Sanghoon
S
Wiersma
Erik J
EJ
D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000821:virology; D005822:Genetic Vectors; D007140:Immunoglobulin Fab Fragments / Q000235:genetics*; D008969:Molecular Sequence Data; D019151:Peptide Library; D014771:Virion / Q000235:genetics
D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library