Mathew P. Robin

Conjugation-induced fluorescent labeling of proteins and polymers using dithiomaleimides.

Dithiomaleimides (DTMs) with alkyl substituents are shown to be a novel class of highly emissive fluorophores. Variable solubility and further functionalization can easily be tailored through the choice of N and S substituents. Inclusion of a DTM unit into a ROP/RAFT initiator or insertion into the disulfide bond of salmon calcitonin (sCT) demonstrates the utility for fluorescent labeling of polymers and proteins. Simultaneous PEGylation and fluorescent labeling of sCT is also demonstrated, using the DTM unit as both a linker and a fluorophore. It is anticipated that DTMs will offer an attractive alternative to commonly used bulky, planar fluorophores.

New functional handle for use as a self-reporting contrast and delivery agent in nanomedicine.

The synthesis and photophysical characterization of a chromophore-bridged block copolymer system is presented. This system is based on a dithiomaleimide (DTM) functional group as a highly emissive functionality which can readily be incorporated into polymeric scaffolds. A key advantage of this new reporter group is its versatile chemistry, ease of further functionalization, and notably small size, which allows for ready incorporation without affecting or disrupting the self-assembly process critical to the formation of core-shell polymeric contrast and drug delivery agents. We demonstrate the potential of this functionality with a diblock system which has been shown to be appropriate for micellization and, when in the micellar state, does not self-quench. The block copolymer is shown to be significantly more emissive than the lone dye, with a concentration-independent emission and anisotropy profile from 1.5 mM to 0.15 μM. An emission lifetime and anisotropy decay comparison of the block copolymer to its micelle displays that time-domain fluorescence lifetime imaging (FLIM) is able to rapidly resolve differences in the supramolecular state of this block-dye-block polymer system. Furthermore, the ability to resolve these differences in the supramolecular state means that the DTM micelles are capable of self-reporting when disassembly occurs, simply by monitoring with FLIM. We demonstrate the great potential for in vitro applications that this system provides by using FLIM to observe micelle disassembly in different vascular components of rat hippocampal tissue. In total this system represents a new class of in-chain emitter which is appropriate for application in quantitative imaging and the tracking of particle degradation/disassembly events in biological environments.

Hollow block copolymer nanoparticles through a spontaneous one-step structural reorganization.

The spontaneous one-step synthesis of hollow nanocages and nanotubes from spherical and cylindrical micelles based on poly(acrylic acid)-b-polylactide (P(AA)-b-P(LA)) block copolymers (BCPs) has been achieved. This structural reorganization, which occurs simply upon drying of the samples, was elucidated by transmission electron microscopy (TEM) and atomic force microscopy (AFM). We show that it was necessary to use stain-free imaging to examine these nanoscale assemblies, as the hollow nature of the particles was obscured by application of a heavy metal stain. Additionally, the internal topology of the P(AA)-b-P(LA) particles could be tuned by manipulating the drying conditions to give solid or compartmentalized structures. Upon resuspension, these reorganized nanoparticles retain their hollow structure and display significantly enhanced loading of a hydrophobic dye compared to the original solid cylinders.

Fluorescent and chemico-fluorescent responsive polymers from dithiomaleimide and dibromomaleimide functional monomers

A new class of brightly fluorescent and profluorescent methacrylate and acrylate monomers is reported. The fluorescent monomers contain the dithiomaleimide (DTM) fluorophore, which imparts a large Stokes shift (up to 250 nm) and bright emission. Furthermore, the simple and efficient chemistry of the DTM group, as well as its excellent processability (highly soluble, neutral functional group) makes monomer preparation straightforward. Copolymerisation at 10 mol% loading with a range of hydrophobic and hydrophilic monomers is demonstrated by RAFT polymerisation. Reactions proceed to high monomer conversion with excellent control over molecular weight (ĐM 95% conversion in 3 h), and by using thiols of different polarities can be used to progressively tune the LCST cloud point of a thermoresponsive polymer over a range of 11 °C. Therefore, both DTM and DBM functional monomers provide a simple and effective tool for fluorescent labelling of (meth)acrylate polymers.

Aminomaleimide fluorophores: a simple functional group with bright, solvent dependent emission

Amino-substituted maleimides form a new class of highly emissive compounds, with large Stokes shifts (>100 nm) and high quantum yields (up to ∼60%).

One-pot synthesis of super-bright fluorescent nanogel contrast agents containing a dithiomaleimide fluorophore

Fluorescent nanogels with radii ranging from 12–17 nm, high quantum yield, concentration-independent emission, and fluorescent lifetimes of ca. 25 ns have been synthesised in a one-pot process. Solutions demonstrate a concentration independent emission between 0.1–10−3 wt%, allowing for true quantitative imaging where dye emission is a measure of local nanoparticle concentration.

Fluorescent polymeric nanovehicles for neural stem cell modulation

Nanomaterials are emerging as strong candidates for applications in drug delivery and offer an alternative platform to modulate the differentiation and activity of neural stem cells. Herein we report the synthesis and characterization of two different classes of polymeric nanoparticles: N-isopropylacrylamide-based thermoresponsive nanogels RM1 and P(TEGA)-b-P(d,lLA)2 nano-micelles RM2. We covalently linked the nanoparticles with fluorescent tags and demonstrate their ability to be internalized and tracked in neural stem cells from the postnatal subventricular zone, without affecting their proliferation, multipotency and differentiation characteristics up to 150 μg ml-1. The difference in chemical structure of RM1 and RM2 does not appear to impact toxicity however it influences the loading capacity. Nanogels RM1 loaded with retinoic acid improve solubility of the drug which is released at 37 °C, resulting in an increase in the number of neurons, comparable to what can be obtained with a solution of the free drug solubilised with a small percentage of DMSO.