Polymeric dipicolylamine based mass tags for mass cytometry

Mass cytometry is an emerging powerful bioanalytical technique for high-dimensional single-cell analysis. In this technique, cells are stained with metal-isotope-tagged antibodies and are analyzed by an inductively coupled plasma time-of-flight mass spectrometer. While there are more than 100 stable isotopes available in the m/z 75 to 209 detection range of the instrument, only about 50 parameters can be measured per cell because current reagents are metal-chelating polymers with pendant aminocarboxylate chelators that only bind hard metal ions such as the rare earths and Bi³⁺. Here we describe the synthesis and characterization of a new type of metal-chelating polymer with pendant dipicolylamine chelators suited to binding intermediate to soft metals such as rhenium and platinum. We introduce two different conjugation strategies, a thiol–maleimide reaction that works well for rhenium, and a DBCO-azide click reaction designed to avoid potential complications of Pt and other heavy metals interacting with thiol groups. We show that these polymers can serve as new elemental mass tags for mass cytometry. Antibody-polymer conjugates of CD20 and CD8a prepared by both coupling reactions were employed in conjunction with commercial metal-conjugated antibodies for multi-parameter single-cell immunoassays.

A new type of metal-chelating polymer with pendant dipicolylamine chelators that bind rhenium and platinum has been developed for mass cytometry applications.     

Effect of oligomers on the polymer diffusion rate in poly(butyl methacrylate) latex films

The aqueous phase of a poly(butyl methacrylate) (PBMA) latex dispersion contained an oligomeric component that was isolated after sedimentation of the PBMA latex particles. The component contained both water‐soluble PBMA oligomer and some longer chain species that were present as a very fine colloidal dispersion. We describe the isolation and characterization of this component. This component was then added to a purified PBMA latex dispersion from which the aqueous component was previously removed. Latex films were prepared, and in the presence of the oligomeric material, the rate of polymer diffusion in the latex film was strongly enhanced. The magnitude of the enhancement was fit quantitatively to the Fujita–Doolittle equation, indicating that the oligomers acted like a traditional plasticizer to increase the free volume in the system. © 2000 John Wiley & Sons, Inc. J Polym Sci A: Polym Chem 38: 3933–3943, 2000     

Non-ionic surfactant effects on polymer diffusion in poly(butyl methacrylate) latex films

The influence of non-ionic surfactant molecules on polymer interdiffusion in poly(butyl methacrylate) (PBMA) latex films was investigated using the non-radiative energy transfer technique. It was found that nonylphenoxypoly(ethylene oxide) surfactants such as NP-20 and NP-100 act as plasticizers to enhance the polymer diffusion rate. The increase in diffusion rate with the increase in surfactant concentration is in complete accord with the Fujita-Doolittle free volume model. This suggests that by the time the latex dispersion is dry, the surfactant has diffused uniformly into the latex polymer.     

Theoretical Study of the Wavelength Selection for the Photocleavage of Coumarin-caged D-luciferin

The equilibrium structures and optical properties of the photolabile caged luciferin, (7-diethylaminocoumarin-4-yl)methyl caged D-luciferin (DEACM-caged D-luciferin), in aqueous solution were investigated via quantum chemical calculations. The probable conformers of DEACM-caged D-luciferin were determined by potential energy curve scans and structural optimizations. We identified 40 possible conformers of DEACM-caged D-luciferin in water by comparing the Gibbs free energy of the optimized structures. Despite the difference in their structures, the conformers were similar in terms of assignments, oscillator strengths and energies of the three low-lying excited states. From the concentrations of the conformers and their oscillator strengths, we obtained a theoretical UV/Vis spectrum of DEACM-caged D-luciferin that has two main bands of shape nearly identical to the experimental UV/Vis spectrum. The absorption bands with maxima ~ 384 and 339 nm were attributed to the electronic excitations of the caged group and the luciferin moiety, respectively, by analysis of the theoretical UV/Vis spectrum. Furthermore, the analysis showed that DEACM-caged D-luciferin is excited in the caged group only by light of wavelength ranging within 400–430 nm, which is in the long-wavelength tail of the 384 nm band. This should be tested to lower damage upon photocleavage.     

Poly(N‐isopropylacrylamide) Phase Diagrams: Fifty Years of Research

In 1968, Heskins and Guillet published the first systematic study of the phase diagram of poly(N‐isopropylacrylamide) (PNIPAM), at the time a “young polymer” first synthesized in 1956. Since then, PNIPAM became the leading member of the growing families of thermoresponsive polymers and of stimuli‐responsive, “smart” polymers in general. Its thermal response is unanimously attributed to its phase behavior. Yet, in spite of 50 years of research, a coherent quantitative picture remains elusive. In this Review we survey the reported phase diagrams, discuss the differences and comment on theoretical ideas regarding their possible origins. We aim to alert the PNIPAM community to open questions in this reputably mature domain.     

Stimulus-responsive self-assembly: reversible, redox-controlled micellization of polyferrocenylsilane diblock copolymers

In depth studies of the use of electron transfer reactions as a means to control the self-assembly of diblock copolymers with an electroactive metalloblock are reported. Specifically, the redox-triggered self-assembly of a series of polystyrene-block-polyferrocenylsilane (PS-b-PFS) diblock copolymers in dichloromethane solution is described. In the case of the amorphous polystyrene(n)-b-poly(ferrocenylphenylmethylsilane)(m) diblock copolymers (PS(n)-b-PFMPS(m): n = 548, m = 73; n = 71, m = 165; where n and m are the number-averaged degrees of polymerization), spherical micelles with an oxidized PFS core and a PS corona were formed upon oxidation of more than 50% of the ferrocenyl units by [N(C(6)H(4)Br-4)(3)][SbX(6)] (X = Cl, F). Analogous block copolymers containing a poly(ferrocenylethylmethylsilane) (PFEMS) metalloblock, which has a lower glass transition temperature, behaved similarly. However, in contrast, on replacement of the amorphous metallopolymer blocks by semicrystalline poly(ferrocenyldimethylsilane) (PFDMS) segments, a change in the observed morphology was detected with the formation of ribbon-like micelles upon oxidation of PS(535)-b-PFDMS(103) above the same threshold value. Again the coronas consisted of fully solvated PS and the core consisted of partially to fully oxidized PFS associated with the counteranions. When oxidation was performed with [N(C(6)H(4)Br-4)(3)][SbF(6)], reduction of the cores of the spherical or ribbon-like micelles with [Co(η-C(5)Me(5))(2)] enabled full recovery of the neutral chains and no significant chain scission was detected.     

Biomimetic hemocompatible coatings through immobilization of hyaluronan derivatives on metal surfaces

Biomimetic coatings offer exciting options to modulate the biocompatibility of biomaterials. The challenge is to create surfaces that undergo specific interactions with cells without promoting nonspecific fouling. This work reports an innovative approach toward biomimetic surfaces based on the covalent immobilization of a carboxylate terminated PEGylated hyaluronan (HA-PEG) onto plasma functionalized NiTi alloy surfaces. The metal substrates were aminated via two different plasma functionalization processes. Hyaluronan, a natural glycosaminoglycan and the major constituent of the extracellular matrix, was grafted to the substrates by reaction of the surface amines with the carboxylic acid terminated PEG spacer using carbodiimide chemistry. The surface modification was monitored at each step by X-ray photoelectron spectroscopy (XPS). HA-immobilized surfaces displayed increased hydrophilicity and reduced fouling, compared to bare surfaces, when exposed to human platelets (PLT) in an in vitro assay with radiolabeled platelets (204.1 +/- 123.8 x 10 (3) PLT/cm (2) vs 538.5 +/- 100.5 x 10 (3) PLT/cm (2) for bare metal, p < 0.05). Using a robust plasma patterning technique, microstructured hyaluronan surfaces were successfully created as demonstrated by XPS chemical imaging. The bioactive surfaces described present unique features, which result from the synergy between the intrinsic biological properties of hyaluronan and the chemical composition and morphology of the polymer layer immobilized on a metal surface.