Chemical Modification of M13 Bacteriophage and Its Application in Cancer Cell Imaging

Abstract

The M13 bacteriophage has been demonstrated to be a robust scaffold for bionanomaterial development. In this paper, we report on the chemical modifications of three kinds of reactive groups, i.e., the amino groups of lysine residues or N-terminal, the carboxylic acid groups of aspartic acid or glutamic acid residues, and the phenol group of tyrosine residues, on M13 surface. The reactivity of each group was identified through conjugation with small fluorescent molecules. Furthermore, the regioselectivity of each reaction was investigated by HPLC-MS-MS. By optimizing the reaction condition, hundreds of fluorescent moieties could be attached to create a highly fluorescent M13 bacteriophage. In addition, cancer cell targeting motifs such as folic acid could also be conjugated onto the M13 surface. Therefore, dual-modified M13 particles with folic acid and fluorescent molecules were synthesized via the selective modification of two kinds of reactive groups. Such dual-modified M13 particles showed very good binding affinity to human KB cancer cells, which demonstrated the potential applications of M13 bacteriophage in bioimaging and drug delivery.

Direct Peptide Bioconjugation/PEGylation at Tyrosine with Linear and Branched Polymeric Diazonium Salts

“…The experiment was then repeated with 15 N labeled (3b)-sCT and (6)-sCT which resulted in the release of sCT with an additional 15 NH2 group (Scheme 5 for (3b)-sCT). Figure 5.…”

“…14 Wang et al have since reported a study into the functionalisation of TMV using a combination of diazonium coupling at tyrosine and and this has then been adapted to modify the tyrosine residues of M13 bacteriophage. 15 Of the three techniques developed by Francis, diazoniummediated targeting of tyrosine residues appeared to us to be be the most attractive for direct polymer conjugation of (poly)peptides. The three-component Mannich-type approach is generally a slow reaction, which makes it less attractive for the coupling of macromolecules, a process that in itself is slower compared to coupling with small molecules.…”

Polymerase Chain Reaction on a Viral Nanoparticle

“…21 Once the pVIII protein was derivatised with SMCC, R-For was then added for conjugation to the M13 via thioether bond formation. The bacteriophage material was then separated from the unreacted components using size exclusion chromatography.…”

Efficient In Vivo Selection of a Novel Tumor-Associated Peptide from a Phage Display Library

“…The minor coat protein pIII that caps the particle is engineered to display five copies of a tumor-avid QFPPKLTNNSML peptide. Five lysine groups (Lys 8, 40, 43, 44, 48) and one N-terminal amino group (Ala 1) on each pVIII subunit are available for chemical modification [28]. (B) Bifunctional p-SCN-Bn-DOTA derivative is added to a 1×10 11 pfu phage in conjugation buffer at pH = 9.…”