T7 displayed peptides as targets for selecting peptide specific scFvs from M13 scFv display libraries.
M13 scFv display libraries are frontline research tools due to the rapidity in which target binding scFv can be obtained. Simple modifications of the standard affinity selection and amplification protocol allow for multiple targets to be panned simultaneously. Target peptide or protein production can become the rate-limiting step in a high throughput approach to antibody selection. To overcome this obstacle and to save both time and money, we have displayed the target peptides on T7 phage particles. In this paper we demonstrate that these particles can be used directly to select peptide specific scFv. We have discovered that the selection is most efficient when a linker separates the display from the T7 coat protein and when a continuous subtraction is employed. We have used antibodies selected in this fashion to characterize target proteins in cell lysates, immuno-precipitations and immuno-histochemistry.
Castillo
J
J
Chiron Corporation, 4560 Horton Street, M/S 4.5, Emeryville, CA 94608, USA.
Goodson
B
B
Winter
J
J
D000918:Antibody Specificity; D017104:Bacteriophage M13 / Q000235:genetics*; D017123:Bacteriophage T7 / Q000235:genetics*; D015153:Blotting, Western; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000821:virology; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D019151:Peptide Library; D010455:Peptides / Q000235:genetics* / Q000276:immunology*
D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D010455:Peptides