Antibodies in haystacks: how selection strategy influences the outcome of selection from molecular diversity libraries.
Antibodies against most antigens can be isolated from high quality phage antibody libraries. However, not all antibodies binding a particular antigen are necessarily found when standard selections are performed. Here we investigate the effect of two different selection strategies on the isolation of antibodies against a number of different antigens, and find that these different strategies tend to select different antibodies, with little overlap between them. This indicates that the full diversity of these libraries is not tapped by a single selection strategy and that each selection strategy imposes different selective criteria in addition to that of antigen binding. To fully exploit such libraries, therefore, many different selection strategies should probably be employed for each antigen. The use of alternative strategies should be considered when selection apparently fails, or when the number of different antibodies recognizing an antigen needs to be maximised. Furthermore, the microtitre selection strategy developed is likely to prove useful in the application of phage antibody libraries to the human genome project, allowing the high throughput selection of antibodies against multiple antigens simultaneously.
Lou
J
J
Department of Anesthesia 3s50, San Francisco General Hospital, UCSF, San Francisco, CA, USA.
Marzari
R
R
Verzillo
V
V
Ferrero
F
F
Pak
D
D
Sheng
M
M
Yang
C
C
Sblattero
D
D
Bradbury
A
A
D000906:Antibodies / Q000235:genetics / Q000276:immunology*; D000916:Antibody Diversity; D000941:Antigens / Q000276:immunology; D002999:Clone Cells / Q000276:immunology; D006801:Humans; D019151:Peptide Library
D000906:Antibodies; D000941:Antigens; D019151:Peptide Library