T7 displayed peptides as targets for selecting peptide specific scFvs fromM13 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 t argets to be panned simultaneously, Target peptide or protein production ca n become the rate-limiting step in a high throughput approach to antibody s election. To overcome this obstacle and to save both time and money, we hav e displayed the target peptides on T7 phage particles. In this paper we dem onstrate that these particles can be used directly to select peptide specif ic scFv. We have discovered that the selection is most efficient when a lin ker separates the display from the T7 coat protein and when a continuous su btraction is employed. We have used antibodies selected in this fashion to characterize target proteins in cell lysates, immuno-precipitations and imm uno-histochemistry. (C) 2001 Elsevier Science B.V. All rights reserved.