Application of phage display to high throughput antibody generation and characterization.
Schofield DJ,
Pope AR,
Clementel V,
Buckell J,
Chapple SDj,
Clarke KF,
Conquer JS,
Crofts AM,
Crowther SR,
Dyson MR,
Flack G,
Griffin GJ,
Hooks Y,
Howat WJ,
Kolb-Kokocinski A,
Kunze S,
Martin CD,
Maslen GL,
Mitchell JN,
O'Sullivan M,
Perera RL,
Roake W,
Shadbolt SP,
Vincent KJ,
Warford A,
Wilson WE,
Xie J,
Young JL,
McCafferty J.
Abcam Ltd, Cambridge Science Park, Cambridge CB4 0FW, UK. Darren.Schofield@abcam.com
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.
PMID: 18047641 [PubMed - indexed for MEDLINE]
PMCID: PMC2258204