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.
Abcam Ltd, Cambridge Science Park, Cambridge CB4 0FW, UK. Darren.Schofield@abcam.com
Darren J
DJ
Schofield
Anthony R
AR
Pope
Veronica
V
Clementel
Jenny
J
Buckell
Susan Dj
SDj
Chapple
Kay F
KF
Clarke
Jennie S
JS
Conquer
Anna M
AM
Crofts
Sandra R E
SR
Crowther
Michael R
MR
Dyson
Gillian
G
Flack
Gareth J
GJ
Griffin
Yvette
Y
Hooks
William J
WJ
Howat
Anja
A
Kolb-Kokocinski
Susan
S
Kunze
Cecile D
CD
Martin
Gareth L
GL
Maslen
Joanne N
JN
Mitchell
Maureen
M
O'Sullivan
Rajika L
RL
Perera
Wendy
W
Roake
S Paul
SP
Shadbolt
Karen J
KJ
Vincent
Anthony
A
Warford
Wendy E
WE
Wilson
Jane
J
Xie
Joyce L
JL
Young
John
J
McCafferty
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