Approaches for Conjugating Tailor-Made Polymers to Proteins

Paeth, Matthew; Stapleton, Jacob; Dougherty, Melissa Lucius; Fischesser, Henry; Shepherd, Jerry T.; McCauley, Matthew; Falatach, Rebecca; Page, Richard C.; Berberich, Jason A.; Konkolewicz, Dominik

doi:10.1016/bs.mie.2016.12.004

Cited by 9 publications

(10 citation statements)

References 66 publications

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“…Horseradish peroxidase type I powder (HRP, 146 units/mg, Sigma, Burlington, VT, USA) was stored at 4 °C. (2-propionic acid)yl ethyl trithiocarbonate (PAETC) was synthesized following procedures described in the literature [49,50].…”

Section: Methodsmentioning

confidence: 99%

Investigating the Mechanism of Horseradish Peroxidase as a RAFT-Initiase

et al. 2018

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A detailed mechanistic and kinetic study of enzymatically initiated RAFT polymerization is performed by combining enzymatic assays and polymerization kinetics analysis. Horseradish peroxidase (HRP) initiated RAFT polymerization of dimethylacrylamide (DMAm) was studied. This polymerization was controlled by 2-(propionic acid)ylethyl trithiocarbonate (PAETC) in the presence of H 2 O 2 as a substrate and acetylacetone (ACAC) as a mediator. In general, well controlled polymers with narrow molecular weight distributions and good agreement between theoretical and measured molecular weights are consistently obtained by this method. Kinetic and enzymatic assay analyses show that HRP loading accelerates the reaction, with a critical concentration of ACAC needed to effectively generate polymerization initiating radicals. The PAETC RAFT agent is required to control the reaction, although the RAFT agent also has an inhibitory effect on enzymatic performance and polymerization. Interestingly, although H 2 O 2 is the substrate for HRP there is an optimal concentration near 1 mM, under the conditions studies, with higher or lower concentrations leading to lower polymerization rates and poorer enzymatic activity. This is explained through a competition between the H 2 O 2 acting as a substrate, but also an inhibitor of HRP at high concentrations.

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Section: Methodsmentioning

confidence: 99%

Investigating the Mechanism of Horseradish Peroxidase as a RAFT-Initiase

et al. 2018

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“…Thus, not only linear homopolymers and random copolymers, but also block and gradient copolymers, and various smart polymers with thermal (lower critical solution temperature (LCST) and upper critical solution temperature (UCST)) or pH or light responsiveness can be grown from proteins . This Perspective will focus on the grafting‐from technique, however, many reports have been published discussing other methods …”

Section: Synthesize: Broadening the Versatility Of Controlled Radicalmentioning

confidence: 99%

Next generation protein‐polymer conjugates

et al. 2018

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“…25,35,36 A number of different chemistries have been used to facilitate grafting of polymers to proteins, including polymers bearing amine-reactive functionality such as NHSesters or anhydrides, 25,36,37 maleimide or dibromomaleimide functionality for reaction with cysteine residues, 25,36,[38][39][40] and bioorthogonal "click" reactions. 25,34,36,41 Grafting-to permits incorporation of both water-soluble and water-insoluble functionalities into the polymer structure. For example, hydrophobic drugs are an important class of water-insoluble molecules that can be incorporated into polymer structures when using the grafting-to approach.…”

Section: Introductionmentioning

confidence: 99%

Protein–Polymer Conjugates Synthesized Using Water-Soluble Azlactone-Functionalized Polymers Enable Receptor-Specific Cellular Uptake toward Targeted Drug Delivery

Kim¹,

Sirois

Cegla³

et al. 2019

Bioconjugate Chem.

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Conjugation of proteins to drug-loaded polymeric structures is an attractive strategy for facilitating target-specific drug delivery for a variety of clinical needs. Polymers currently available for conjugation to proteins generally have limited chemical versatility for subsequent drug loading. Many polymers that do have chemical functionality useful for drug loading are often insoluble in water, making it difficult to synthesize functional protein-polymer conjugates for targeted drug delivery. In this work, we demonstrate that reactive, azlactone-functionalized polymers can be grafted to proteins, conjugated to a small-molecule fluorophore, and subsequently internalized into cells in a receptor-specific manner. Poly(2-vinyl-4,4-dimethylazlactone), synthesized using reversible addition-fragmentation chain transfer polymerization, was modified postpolymerization with substoichiometric equivalents of triethylene glycol monomethyl ether to yield reactive water-soluble, azlactone-functionalized copolymers. These reactive polymers were then conjugated to proteins holo-transferrin and ovotransferrin. Protein gel analysis verified successful conjugation of proteins to polymer, and protein-polymer conjugates were subsequently purified from unreacted proteins and polymers using size exclusion chromatography. Internalization experiments using a breast cancer cell line that overexpresses the transferrin receptor on its surface showed that the holo-transferrin-polymer conjugate was successfully internalized by cells in a

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Approaches for Conjugating Tailor-Made Polymers to Proteins

Cited by 9 publications

References 66 publications

Investigating the Mechanism of Horseradish Peroxidase as a RAFT-Initiase

Investigating the Mechanism of Horseradish Peroxidase as a RAFT-Initiase

Next generation protein‐polymer conjugates

Protein–Polymer Conjugates Synthesized Using Water-Soluble Azlactone-Functionalized Polymers Enable Receptor-Specific Cellular Uptake toward Targeted Drug Delivery

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