scholarly journals An In-Depth Analysis Approach Enabling Precision Single Chain Nanoparticle Design

2020 ◽  
Author(s):  
Johanna Engelke ◽  
Bryan T. Tuten ◽  
Ralf Schweins ◽  
Hartmut Komber ◽  
Leonie Barner ◽  
...  
Keyword(s):  
Molar Mass ◽  
Step Change ◽  
Unique Combination ◽  
Single Chain ◽  
Reactive Group ◽  
Depth Analysis ◽  
Precursor Polymers ◽  
Nanoparticle Design ◽  
Group Density ◽  
A Minor

The formation of single chain nanoparticles (SCNPs) is typically characterized by a shift towards higher elution volumes in SEC due to size decrease upon folding. However, a step-change in SCNP analysis is required for understanding of the nature of intramolecular SCNP folding. Herein, we exploit a unique combination of SANS, 19F NMR spectroscopy, and multidetection SEC to generate a systematic view of the folded morphology of poly(butyl acrylate) based-SCNPs as a function of their reactive group density and molar mass. In addition to detailed morphological insights, we establish that the primary factor dictating the compaction of SCNPs is their reactive group density, which is ineffective below 5 mol%, reaching maximum compaction close to 30 mol%. Above 20 kDa the molar mass of the precursor polymers has a minor impact on how an SCNP compacts.

scholarly journals An In-Depth Analysis Approach Enabling Precision Single Chain Nanoparticle Design

2020 ◽  
Author(s):  
Johanna Engelke ◽  
Bryan T. Tuten ◽  
Ralf Schweins ◽  
Hartmut Komber ◽  
Leonie Barner ◽  
...  
Keyword(s):  
Molar Mass ◽  
Step Change ◽  
Unique Combination ◽  
Single Chain ◽  
Reactive Group ◽  
Depth Analysis ◽  
Precursor Polymers ◽  
Nanoparticle Design ◽  
Group Density ◽  
A Minor

The formation of single chain nanoparticles (SCNPs) is typically characterized by a shift towards higher elution volumes in SEC due to size decrease upon folding. However, a step-change in SCNP analysis is required for understanding of the nature of intramolecular SCNP folding. Herein, we exploit a unique combination of SANS, 19F NMR spectroscopy, and multidetection SEC to generate a systematic view of the folded morphology of poly(butyl acrylate) based-SCNPs as a function of their reactive group density and molar mass. In addition to detailed morphological insights, we establish that the primary factor dictating the compaction of SCNPs is their reactive group density, which is ineffective below 5 mol%, reaching maximum compaction close to 30 mol%. Above 20 kDa the molar mass of the precursor polymers has a minor impact on how an SCNP compacts.

scholarly journals An in-depth analysis approach enabling precision single chain nanoparticle design

2020 ◽  
Vol 11 (41) ◽  
pp. 6559-6578 ◽  
Author(s):  
Johanna Engelke ◽  
Bryan T. Tuten ◽  
Ralf Schweins ◽  
Hartmut Komber ◽  
Leonie Barner ◽  
...  
Keyword(s):  
Step Change ◽  
Analysis Approach ◽  
Single Chain ◽  
Depth Analysis ◽  
Nanoparticle Design ◽  
Polymer Folding

The synthesis of single chain nanoparticles (SCNPs) is a vibrant field in macromolecular science. However, to achieve an in-depth understanding of the nature of intramolecular polymer folding, a step-change in the methodologies for SCNP analysis is required.

Demonstration of a Functionally Active tPA-Like Plasminogen Activator in Human Platelets

1994 ◽  
Vol 71 (04) ◽  
pp. 493-498 ◽  
Author(s):  
D L Wang ◽  
Y T Pan ◽  
J J Wang ◽  
C H Cheng ◽  
C Y Liu
Keyword(s):  
Plasminogen Activator ◽  
Fibrinolytic Activity ◽  
Protein Band ◽  
Human Platelets ◽  
Free Form ◽  
Plasminogen Activation ◽  
Single Chain ◽  
Western Blots ◽  
A Minor ◽  
Pai 1

SummaryThe mechanism of platelet-enhanced fibrinolysis is unclear. We therefore investigated the fibrinolytic activity of human platelets and demonstrated that they contain a tissue plasminogen activator (tPA)- like plasminogen activator, abbreviated as tPA-like-PA. This activator was detected by ELISA in platelet incubation medium and in platelet Triton extracts. Plasminogen activation assays showed that this tPA- like-PA could induce plasminogen activation to form plasmin. Western blots of Triton extracts incubated with anti-tPA antibody demonstrated a major 64-kD protein band, compared to a 70-kD band for standard single chain tPA, plus a minor 118-kD band corresponding to a complex of tPA-like-PA and plasminogen activator inhibitor (PAI-1). Western blots of Triton extracts incubated with anti-PAI-1 antibody produced an approximately similar high-molecular-weight (118 kD) protein band. Fibrin zymographic analysis of affinity-purified tPA-like- PA demonstrated a major and a minor fibrin lysis zone, which approximately corresponded to the tPA-like-PA and its complex with PAI-1 observed by Western blots. Immunogold labelling and electron microscopy demonstrated that platelet activator, either as the free form or co-localized with PAI-1, was present in granules and in channels of the open canalicular system. We conclude that platelets contain a functionally active tPA-like-PA, whose low fibrinolytic activity might be due to its readily forming a complex with PAI-1. This functionally active tPA-like-PA might contribute to the enhanced fibrinolytic activity of platelets observed in platelet-rich thrombi.

Tuning the magnetization dynamics of TbIII-based single-chain magnets through substitution on the nitronyl nitroxide radical

2019 ◽  
Vol 48 (24) ◽  
pp. 8989-8994 ◽  
Author(s):  
Xiaoqing Liu ◽  
Yu-Xia Wang ◽  
Zongsu Han ◽  
Tian Han ◽  
Wei Shi ◽  
...  
Keyword(s):  
Nitroxide Radical ◽  
Nitronyl Nitroxide ◽  
Single Chain ◽  
Magnetization Dynamics ◽  
Nitronyl Nitroxide Radical ◽  
Magnetic Couplings ◽  
A Minor ◽  
Single Chain Magnets

A minor change of the site of the substituent on the nitronyl nitroxide radical significantly influences the magnetization dynamics of two single-chain magnets, due to the influence of both the magnetic anisotropies of the TbIII centers and magnetic couplings along the chains.

scholarly journals Production, purification and characterization of the catalytic domain of glucoamylase from Aspergillus niger

1993 ◽  
Vol 292 (1) ◽  
pp. 197-202 ◽  
Author(s):  
B Stoffer ◽  
T P Frandsen ◽  
P K Busk ◽  
P Schneider ◽  
I Svendsen ◽  
...  
Keyword(s):  
Aspergillus Niger ◽  
Catalytic Domain ◽  
Limited Proteolysis ◽  
Exchange Chromatography ◽  
High Yield ◽  
Amino Acid Residues ◽  
Single Chain ◽  
Subtilisin Carlsberg ◽  
A Minor

The catalytic domain of glucoamylases G1 and G2 from Aspergillus niger is produced in vitro in high yield by limited proteolysis using either subtilisin Novo or subtilisin Carlsberg. Purification by affinity chromatography on an acarbose-Sepharose column followed by ion-exchange chromatography on HiLoad Q-Sepharose leads to separation of a number of structurally closely related forms of domain. The cleavage occurs primarily between Val-470 and Ala-471 as indicated by C-terminal sequencing, whereas the N-terminus is intact. Subtilisin Carlsberg, in addition, produces a type of domain which is hydrolysed before Ser-444, an O-glycosylated residue. This leaves the fragment Ser-444-Val-470 disulphide-bonded to the large N-terminal part of the catalytic domain. Subtilisin Novo, in contrast, tends to yield a minor fraction of forms extending approx. 30-40 amino-acid residues beyond Val-470. The thermostability is essentially the same for the single-chain catalytic domain and the original glucoamylases G1 and G2, whereas the catalytic domain cut between Ser-443 and Ser-444 is less thermostable. For both types of domain the kinetic parameters, Km and kcat., for hydrolysis of maltose are very close to the values found for glucoamylases G1 and G2.

scholarly journals The ecogenomics of dsDNA bacteriophages in feces of stabled and feral horses

2020 ◽  
Author(s):  
V. V. Babenko ◽  
A. Millard ◽  
E. E. Kulikov ◽  
N.N. Spasskaya ◽  
M. A. Letarova ◽  
...  
Keyword(s):  
Animal Species ◽  
Equus Caballus ◽  
Isolated Populations ◽  
Feral Horses ◽  
Genus Level ◽  
Minor Fraction ◽  
Depth Analysis ◽  
Ecological Parameters ◽  
A Minor ◽  
Geographically Isolated

AbstractThe viromes of the mammalian lower gut were shown to be heavily dominated by bacteriophages; however, only for humans were the composition and intervariability of the bacteriophage communities studied in depth. Here we present an ecogenomics survey of dsDNA bacteriophage diversity in the feces of horses (Equus caballus), comparing two groups of stabled horses, and a further group of feral horses that were isolated on an island. Our results indicate that the dsDNA viromes of the horse feces feature higher richness than in human viromes, with more even distribution of genotypes. No over-represented phage genotypes, such as CrAssphage-related viruses found in humans, were identified. Additionally, many bacteriophage genus-level clusters were found to be present in all three geographically isolated populations. The diversity of the horse intestinal bacteriophages is severely undersampled, and so consequently only a minor fraction of the phage contigs could be linked with the bacteriophage genomes. Our study indicates that bacteriophage ecological parameters in the intestinal ecosystems in horses and humans differ significantly, leading them to shape their corresponding viromes in different ways. Therefore, the diversity and structure of the intestinal virome in different animal species needs to be experimentally studied.Short abstract (needed in some journals as eLife)The viromes of the mammalian gut were shown to be heavily dominated by bacteriophages; however, only for humans were the composition and intervariability of the bacteriophage communities studied in depth. Here we present an ecogenomics survey of dsDNA bacteriophage diversity in the feces of horses (Equus caballus), comparing stabled horses, and feral horses that were isolated on an island. The viromes equine fecal viromes feature higher richness than in human viromes, with more even distribution of genotypes. No over-represented phage genotypes were identified. Additionally, many bacteriophage genus-level clusters were found to be present in geographically isolated populations. Only a minor fraction of the phage contigs could be linked with the bacteriophage genomes. Our study indicates that bacteriophage ecological parameters in the intestinal ecosystems in horses and humans differ significantly, leading them to shape their corresponding viromes in different ways.Importance. (needed for mBio)The study presents the first in depth analysis of the composition and variability of the gut dsDNA bacteriophage community in the mammalian species, other than humans. The study demonstrates that the bacteriophage ecology in the gut is substantially different in different animal species. The results also indicate that the genetic diversity of the equine intestinal bacteriophages is immense and almost totally unexplored by the moment.

Photochemistry in Confined Environments for Single-Chain Nanoparticle Design

2018 ◽  
Vol 140 (30) ◽  
pp. 9551-9557 ◽  
Author(s):  
Hendrik Frisch ◽  
Jan P. Menzel ◽  
Fabian R. Bloesser ◽  
David E. Marschner ◽  
Kai Mundsinger ◽  
...  
Keyword(s):  
Single Chain ◽  
Nanoparticle Design

In vivo Production of Functional Single-Chain Fv Fragment with an N-Terminal-Specific Bio-orthogonal Reactive Group

ChemBioChem ◽  
2010 ◽  
Vol 11 (4) ◽  
pp. 498-501 ◽  
Author(s):  
Edwardraja Selvakumar ◽  
Neelamegam Rameshkumar ◽  
Sun-Gu Lee ◽  
Soo-Jae Lee ◽  
Hyung-Soon Park
Keyword(s):  
Single Chain ◽  
Reactive Group ◽  
Single Chain Fv ◽  
Single Chain Fv Fragment

scholarly journals Accumulation of Unfavorable Biopsychological and Sociocultural Factors in the Course of the Demoralization Process a Minor with a Sensory Hearing Impairment

2021 ◽  
Vol 40 (3) ◽  
pp. 199-217
Author(s):  
Jolanta Bożena Lipińska-Lokś
Keyword(s):  
Risk Factors ◽  
Protective Factors ◽  
Environmental Conditions ◽  
Sociocultural Factors ◽  
Social Maladjustment ◽  
Social Situations ◽  
Low Degree ◽  
Depth Analysis ◽  
A Minor ◽  
New Phenomena

The reality is full of new phenomena, and the existing ones are transformed. An example is the phenomenon of social maladjustment. There is an increase in the number of criminal acts and acts proving demoralization committed by children and adolescents. The situation forces us to take radical measures, preceded by an in-depth analysis of the mechanisms and causes of the problem. The analysis shows the diversification of the demoralization phenomenon and, most often, the accumulation of risk factors for the emergence of the problem of social maladjustment, with the scarcity of the so-called protective factors. People with disabilities are in a particularly disadvantaged situation, often with a low degree of adaptation, exposed to unfavorable social situations. The greater the danger, the less favorable the environmental conditions - family and lack of support.

scholarly journals High-throughput retrieval of physical DNA for NGS-identifiable clones in phage display library

2018 ◽  
Author(s):  
Jinsung Noh ◽  
Okju Kim ◽  
Yushin Jung ◽  
Haejun Han ◽  
Jung-Eun Kim ◽  
...  
Keyword(s):  
High Throughput ◽  
Antibody Fragment ◽  
Phage Display Library ◽  
Sequence Information ◽  
Single Chain ◽  
Depth Analysis ◽  
Scfv Library ◽  
Linkage Information ◽  
Antibody Discovery ◽  
Human Hepatocyte Growth Factor

AbstractIn antibody discovery, in-depth analysis of an antibody library and high-throughput retrieval of clones in the library are crucial to identifying and exploiting rare clones with different properties. However, existing methods have several technical limitations such as low process throughput from laborious cloning process and waste of the phenotypic screening capacity from unnecessary repetitive tests on the dominant clones. To overcome the limitations, we developed a new high-throughput platform for the identification and retrieval of clones in the library, TrueRepertoire™. TrueRepertoire™ provides highly accurate sequences of the clones with linkage information between heavy and light chains of the antibody fragment. Additionally, the physical DNA of clones can be retrieved in high throughput based on the sequence information. We validated the high accuracy of the sequences and demonstrated that there is no platform-specific bias. Moreover, the applicability of TrueRepertoire™ was demonstrated by a phage-displayed single-chain variable fragment (scFv) library targeting human hepatocyte growth factor (hHGF) protein.

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