Phage Wrapping with Cationic Polymers Eliminates Nonspecific Binding between M13 Phage and High p<i>I</i> Target Proteins

Lamboy, Jorge A.; Arter, Jessica A.; Knopp, Kristeene A.; Der, Denise; Overstreet, Cathie M.; Palermo, Edmund F.; Urakami, Hiromitsu; Yu, Ting; Tezgel, Ozgul; Tew, Gregory N.; Guan, Zhibin; Kuroda, Kenichi; Weiss, Gregory A.

doi:10.1021/ja9050873

Cited by 25 publications

(22 citation statements)

References 18 publications

(34 reference statements)

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“…Synthesis of PEDOT nanowires requires LiClO 4 dissolved in a monomeric, aqueous solution of EDOT, and the ClO 4 - anions are closely associated with the PEDOT nanowires during the oxidative electrodeposition. Our strategy takes advantage of the overall highly negatively charged surface of the phage, a property exploited previously for the coating of phage with cationic polymers 51. Thus, the viruses competed with ClO 4 - ions for electrostatic incorporation into the nanowires (Scheme 1b), and remained stable through multiple steps including drying, washing, and treatment with both acetone and nitric acid (0.8 M).…”

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confidence: 99%

Virus-PEDOT Nanowires for Biosensing

et al. 2010

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The separate fields of conducting polymer-based electrochemical sensors and virus-based molecular recognition offer numerous advantages for biosensing. Grafting M13 bacteriophage into an array of poly (3,4-ethylenedioxythiophene) (PEDOT) nanowires generated hybrids of conducting polymers and viruses. The virus incorporation into the polymeric backbone of PEDOT occurs during electropolymerization via lithographically patterned nanowire electrodeposition (LPNE). The resultant arrays of virus-PEDOT nanowires enable real-time, reagent-free electrochemical biosensing of analytes in physiologically relevant buffers.

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confidence: 99%

Virus-PEDOT Nanowires for Biosensing

et al. 2010

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“…We attribute this to nonspecific binding of the positive PEI-QDs to the M13 phages which carry a high negative surface charge. 50 The lack of GSH43-scFv binding to the negative charged Inv-QD is also in contrast to the phage format results (Figure 2c). We attribute this to differences in binding affinity, as from dot blot concentration studies we did observe strong binding of the GSH-scFv to the Inv-QD with increasing GSH-scFv concentration (5–20 μg/mL) with a continued absence of binding to PEI-QDs (Supplementary Figure S11).…”

Section: Resultsmentioning

confidence: 78%

Development and characterization of antibody reagents for detecting nanoparticles

et al. 2015

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The increasing use of nanoparticles (NPs) in technological applications and in commercial products has escalated environmental health and safety concerns. The detection of NPs in the environment and in biological systems is challenged by limitations associated with commonly used analytical techniques. In this paper we report on the development and characterization of NP binding antibodies, termed NProbes. Phage display methodology was used to discover antibodies that bind NPs dispersed in solution. We present a proof-of-concept for the generation of NProbes and their use for detecting quantum dots and titanium dioxide NPs in vitro and in an ex vivo human skin model. Continued development and refinement of NProbes to detect NPs that vary in composition, shape, size, and surface coating will comprise a powerful tool kit that can be used to advance nanotechnology research particularly in the nanotoxicology and nanotherapeutics fields.

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“…This collection of peptide sequences included 22 different configurations of disulfide-constrained peptide libraries and one linear peptide library, ranging in length from 8- to 20-mers 37,38 (Table S1 in Supporting Information).…”

Section: Resultsmentioning

confidence: 99%

Hydrazide Reactive Peptide Tags for Site-Specific Protein Labeling

Eldridge

Weiss

2011

Bioconjugate Chem.

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New site-specific protein labeling (SSPL) reactions for targeting specific, short peptides could be useful for the real time detection of proteins inside of living cells. One SSPL approach matches bioorthogonal reagents with complementary peptides. Here, hydrazide reactive peptides were selected from phage-displayed libraries using reaction-based selections. Selection conditions included washes of varying pH and treatment with NaCNBH3 in order to specifically select reactive carbonyl containing peptides. Selected peptides were fused to T4 lysozyme or synthesized on filter paper for colorimetric assays of the peptide-hydrazide interaction. A peptide-lysozyme protein fusion demonstrated specific, covalent labeling by the Hydrazide Reactive (HyRe) peptides in crude bacterial cell lysates, sufficient for the specific detection of an over-expressed protein fusion. Chemical synthesis of a short HyRe tag variant and subsequent reaction with two structurally distinct hydrazide probes produced covalent adducts observable by MALDI-TOF MS and MS/MS. Rather than isolating reactive carbonyl-containing peptides, we observed reaction with the N–terminal His of HyRe tag 114, amino acid sequence HKSNHSSKNRE, which attacks the hydrazide carbonyl at neutral pH. However, at the pH used during selection wash steps (<6.0), an alternative imine-containing product is formed that can be reduced with sodium cyanoborohydride. MSMS further reveals that this low pH product forms an adduct on Ser6. Further optimization of the novel bimolecular reaction described here could provide a useful tool for in vivo protein labeling and bioconjugate synthesis. The reported selection and screening methods could be widely applicable to the identification of peptides capable of other site-specific protein labeling reactions with bioorthogonal reagents.

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Phage Wrapping with Cationic Polymers Eliminates Nonspecific Binding between M13 Phage and High pI Target Proteins

Cited by 25 publications

References 18 publications

Virus-PEDOT Nanowires for Biosensing

Virus-PEDOT Nanowires for Biosensing

Development and characterization of antibody reagents for detecting nanoparticles

Hydrazide Reactive Peptide Tags for Site-Specific Protein Labeling

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