High-throughput identification of phage-derived imaging agents.
The use of phage-displayed peptide libraries is a powerful method for selecting peptides with desired binding properties. However, the validation and prioritization of "hits" obtained from this screening approach remains challenging. Here, we describe the development and testing of a new analysis method to identify and display hits from phage-display experiments and high-throughput enzyme-linked immunosorbent assay screens. We test the method using a phage screen against activated macrophages to develop imaging agents with higher specificity for active disease processes. The new methodology should be useful in identifying phage hits and is extendable to other library screening methods such as small-molecule and nanoparticle libraries.
Kelly
Kimberly A
KA
Massachusetts General Hospital and Harvard Medical School, Charlestown 02129, USA. kkelly9@partners.org
Clemons
Paul A
PA
Yu
Amy M
AM
Weissleder
Ralph
R
D000345:Affinity Labels / Q000737:chemistry; D001435:Bacteriophages / Q000737:chemistry* / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009000:Monocytes / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D020298:U937 Cells
D000345:Affinity Labels; D008070:Lipopolysaccharides; D019151:Peptide Library; D010455:Peptides