N-Terminal labeling of filamentous phage to create cancer marker imaging agents
| Autor: | Douglas S. Clark, Zachary M. Carrico, Sonny C. Hsiao, Michelle E. Farkas, Matthew B. Francis, Harshal A. Chokhawala, James D. Marks, Yu Zhou |
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| Rok vydání: | 2012 |
| Předmět: |
bioorthogonal
Phage display bloconjugation materials science viruses General Physics and Astronomy Contrast Media Breast Neoplasms Biology Article Fluorescence Cell Line cancer imaging chemistry.chemical_compound Inovirus Epidermal growth factor Cell Line Tumor Breast Cancer Biomarkers Tumor Humans General Materials Science Nanoscience & Nanotechnology Multiphoton Cancer chemistry.chemical_classification Microscopy Bioconjugation Tumor Staining and Labeling General Engineering Amino acid Molecular Imaging Microscopy Fluorescence Multiphoton chemistry Capsid Biochemistry Cell culture Capsid Proteins Bioorthogonal chemistry phage display Ethylene glycol Biomarkers Biotechnology |
| Zdroj: | ACS nano, vol 6, iss 8 |
| Popis: | We report a convenient new technique for the labeling of filamentous phage capsid proteins. Previous reports have shown that phage coat protein residues can be modified, but the lack of chemically distinct amino acids in the coat protein sequences makes it difficult to attach high levels of synthetic molecules without altering the binding capabilities of the phage. To modify the phage with polymer chains, imaging groups, and other molecules, we have developed chemistry to convert the N-terminal amines of the ~4,200 coat proteins into ketone groups. These sites can then serve as chemospecific handles for the attachment of alkoxyamine groups through oxime formation. Specifically, we demonstrate the attachment of fluorophores and up to 3,000 molecules of 2 kD poly(ethylene glycol) (PEG2k) to each of the phage capsids without significantly affecting the binding of phage-displayed antibody fragments to EGFR and HER2 (two important epidermal growth factor receptors). We also demonstrate the utility of the modified phage for the characterization of breast cancer cells using multicolor fluorescence microscopy. Due to the widespread use of filamentous phage as display platforms for peptide and protein evolution, we envision that the ability to attach large numbers of synthetic functional groups to their coat proteins will be of significant value to the biological and materials communities. |
| Databáze: | OpenAIRE |
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