4 research outputs found
Structural transition of nanogel star polymers with pH by controlling PEGMA interactions with acid or base copolymers
<p>We use small angle X-ray scattering (SAXS) to characterise a class of star diblock polymers with a nanogel core on which the outer block arms are comprised of random copolymers of temperature sensitive PEGMA with pH sensitive basic (PDMAEMA) and acidic (PMAA) monomers. The acquired SAXS data show that many of the nanogel star polymers undergo a sharp structural transition over a narrow range of pH, but with unexpectedly large shifts in the apparent pKa with respect to that of the acidic or basic monomer unit, the linear polymer form or even an alternate star polymer with a tightly cross-linked core chemistry. We have demonstrated a distinct and quantifiable structural response for the nanogel star copolymers by altering the core or by pairing the monomers PDMAEMA–PEGMA and PMAA–PEGMA to achieve structural transitions that have typically been observed in stars through changes in arm length and number.</p> <p></p
Recommended from our members
Lipid Cross-Linking of Nanolipoprotein Particles Substantially Enhances Serum Stability and Cellular Uptake
Nanolipoprotein
particles (NLPs) consist of a discoidal phospholipid
lipid bilayer confined by an apolipoprotein belt. NLPs are a promising
platform for a variety of biomedical applications due to their biocompatibility,
size, definable composition, and amphipathic characteristics. However,
poor serum stability hampers the use of NLPs for in vivo applications
such as drug formulation. In this study, NLP stability was enhanced
upon the incorporation and subsequent UV-mediated intermolecular cross-linking
of photoactive DiynePC phospholipids in the lipid bilayer, forming
cross-linked nanoparticles (X-NLPs). Both the concentration of DiynePC
in the bilayer and UV exposure time significantly affected the resulting
X-NLP stability in 100% serum, as assessed by size exclusion chromatography
(SEC) of fluorescently labeled particles. Cross-linking did not significantly
impact the size of X-NLPs as determined by dynamic light scattering
and SEC. X-NLPs had essentially no degradation over 48 h in 100% serum,
which is a drastic improvement compared to non-cross-linked NLPs (50%
degradation by ∼10 min). X-NLPs had greater uptake into the
human ATCC 5637 bladder cancer cell line compared to non-cross-linked
particles, indicating their potential utility for targeted drug delivery.
X-NLPs also exhibited enhanced stability following intravenous administration
in mice. These results collectively support the potential utility
of X-NLPs for a variety of in vivo applications
Process Robustness in Lipid Nanoparticle Production: A Comparison of Microfluidic and Turbulent Jet Mixing
The recent clinical and commercial success of lipid nanoparticles
(LNPs) for nucleic acid delivery has incentivized the development
of new technologies to manufacture LNPs. As new technologies emerge,
researchers must determine which technologies to assess and how to
perform comparative evaluations. In this article, we use a quality-by-design
approach to systematically investigate how the mixer technology used
to form LNPs influences LNPstructure. Specifically, a coaxial turbulent
jet mixer and a staggered herringbone microfluidic mixer were systematically
compared via matched formulation and process conditions. A full-factorial
design-of-experiments study with three factors and three levels was
executed for each mixer to compare process robustness in the production
of antisense oligonucleotide (ASO) LNPs. ASO-LNPs generated with the
coaxial turbulent jet mixer were consistently smaller, had a narrower
particle size distribution, and had a higher ASO encapsulation as
compared to the microfluidic mixer, but had a greater variation in
internal structure with less ordered cores. A subset of the study
was replicated for mRNA-LNPs with comparable trends in particle size
and encapsulation, but more frequent bleb features for LNPs produced
by the coaxial turbulent jet mixer. The study design used here provides
a road map for how researchers may compare different mixer technologies
(or process changes more broadly) and how such studies can inform
process robustness and manufacturing control strategies
Structural and Functional Characterization of CalS11, a TDP-Rhamnose 3′‑<i>O</i>‑Methyltransferase Involved in Calicheamicin Biosynthesis
Sugar
methyltransferases (MTs) are an important class of tailoring
enzymes that catalyze the transfer of a methyl group from <i>S</i>-adenosyl-l-methionine to sugar-based <i>N</i>-, <i>C</i>- and <i>O</i>-nucleophiles.
While sugar <i>N</i>- and <i>C</i>-MTs involved
in natural product biosynthesis have been found to act on sugar nucleotide
substrates prior to a subsequent glycosyltransferase reaction, corresponding
sugar <i>O</i>-methylation reactions studied thus far occur
after the glycosyltransfer reaction. Herein we report the first <i>in vitro</i> characterization using <sup>1</sup>H–<sup>13</sup>C-gHSQC with isotopically labeled substrates and the X-ray
structure determination at 1.55 Å resolution of the TDP-3′-<i>O</i>-rhamnose-methyltransferase CalS11 from <i>Micromonospora
echinospora</i>. This study highlights a unique NMR-based methyltransferase
assay, implicates CalS11 to be a metal- and general acid/base-dependent <i>O</i>-methyltransferase, and as a first crystal structure for
a TDP-hexose-<i>O</i>-methyltransferase, presents a new
template for mechanistic studies and/or engineering