Application of phage display to high throughput antibody generation and characterization.
We have created a high quality phage display library containing over 1010 human antibodies and describe its use in the generation of antibodies on an unprecedented scale. We have selected, screened and sequenced over 38,000 recombinant antibodies to 292 antigens, yielding over 7,200 unique clones. 4,400 antibodies were characterized by specificity testing and detailed sequence analysis and the data/clones are available online. Sensitive detection was demonstrated in a bead based flow cytometry assay. Furthermore, positive staining by immunohistochemistry on tissue microarrays was found for 37% (143/381) of antibodies. Thus, we have demonstrated the potential of and illuminated the issues associated with genome-wide monoclonal antibody generation.
Schofield
Darren J
DJ
Abcam Ltd, Cambridge Science Park, Cambridge CB4 0FW, UK. Darren.Schofield@abcam.com
Pope
Anthony R
AR
Clementel
Veronica
V
Buckell
Jenny
J
Chapple
Susan Dj
SDj
Clarke
Kay F
KF
Conquer
Jennie S
JS
Crofts
Anna M
AM
Crowther
Sandra R E
SR
Dyson
Michael R
MR
Flack
Gillian
G
Griffin
Gareth J
GJ
Hooks
Yvette
Y
Howat
William J
WJ
Kolb-Kokocinski
Anja
A
Kunze
Susan
S
Martin
Cecile D
CD
Maslen
Gareth L
GL
Mitchell
Joanne N
JN
O'Sullivan
Maureen
M
Perera
Rajika L
RL
Roake
Wendy
W
Shadbolt
S Paul
SP
Vincent
Karen J
KJ
Warford
Anthony
A
Wilson
Wendy E
WE
Xie
Jane
J
Young
Joyce L
JL
McCafferty
John
J
D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051379:Mice; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
D017931:DNA Primers; D011994:Recombinant Proteins