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ATLAS scFV Library

ATLAS scFV Library

To enhance our proteomic resources Source BioScience are now distributing the ATLAS clones constructed and annotated at the Wellcome Trust Sanger Institute by team 86.

Antibodies were selected against 404 antigen targets which represent 280 mouse genes. The antigens were primarily produced in E.coli or within mammalian expression system; the majority of these genes are cell surface receptors.

The scFv constructs are fused to a tri-FLAG tag sequence and the dimeric bacterial enzyme, alkaline phosphatase. This has a dual rationale of driving dimerisation which increases avidity of binding and also acts as a useful enzymatic tag for detection purposes. 22 unique clones were selected for each antigen. Antibodies were characterised by specificity testing and detailed sequence analysis. Furthermore, many of scFvs have been used in immunohistochemistry analysis on tissue microarrays.

http://genomebiology.biomedcentral.com/articles/10.1186/gb-2007-8-11-r254

Source BioScience offers the complete clone set of 53 96-well microtitre plates or individual clones, for research purposes only. For each antigen, 22 unique scFv constructs are available.

Protocols / Clone Handling

Plates contain LB broth with 8% glycerol and the appropriate antibiotic. The plates are sent on Dry Ice, and should be stored at -70°C.

Individual clones have been streaked onto LB agar containing 30μl/ml Kanamycin. Please store them at 4°C (not in a freezer). As with all clones, it is recommended that you re-streak to obtain several single colonies to test. As soon as possible, transfer the culture to LB broth containing the antibiotic +8% glycerol and incubate overnight at 30°C (do not try to transfer single colonies, instead scoop out the whole bacterial culture using a disposable inoculation loop). Incubate overnight at 37°C for subsequent freezing at -70°C.

For further details on this library and using the clones please see the publications below:

Application of phage display to high throughput antibody generation and characterization.

A simple vector system to improve performance and utilisation of recombinant antibodies

References

Application of phage display to high throughput antibody generation and characterization.
Schofield DJ, Pope AR, Clementel V, Buckell J, Chapple SD, 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.
Genome Biol. 2007 Nov 29;8(11):R254 [Epub ahead of print]
The Wellcome Trust Sanger Institute, Hixton Campus, Cambridge UK


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