scholarly journals Time grid-based isomer specific N-glycan analysis and detection of bisecting Lewis X in human brain

2021 ◽  
Author(s):  
Johannes Helm ◽  
Clemens Gruenwald-Gruber ◽  
Andreas Thader ◽  
Jonathan Urteil ◽  
Johannes Fuehrer ◽  
...  
Keyword(s):  
Human Brain ◽  
Retention Time ◽  
Protein Glycosylation ◽  
Lewis X ◽  
Structural Assignment ◽  
Novel Approach ◽  
Time Determination ◽  
Isomeric Structures ◽  
Glycan Type ◽  
Time Grid

The importance of protein glycosylation in the biomedical field demands for methods capable of resolving and identifying isomeric structures of N-glycans. However, the unambiguous identification of isomeric structures from complex mixtures is currently not reasonably realized even by the most sophisticated approaches. Here we present a novel approach which uses stable isotope labelled reference N-glycans to establish a retention time grid (glyco-TiGr) on porous graphitized carbon. This furthermore enables retention as the primary criterion for the structural assignment of isomeric N-glycans. Moreover, we biosynthesized forty natural isomers of the fundamental N-glycan type consisting of five hexoses, four N-acetylhexosamines and one fucose residue. Nearly all of these isomers occupied unique positions on the retention time grid. Reference glycan assisted retention time determination with deci-minute accuracy narrowed the assignment space to very few, often only one possible glycan isomer. Application of the glyco-TiGr approach revealed yet undescribed isomers of Lewis x determinants in multimeric human IgA and hybrid type N-glycans in human brain with galactose and even fucose linked to the bisecting N-acetylglucosamine. Thus, the brain N-glycome displayed a degree of sophistication commensurate with this organ's role.

scholarly journals Fabrication of Carbohydrate Chips Based on Polydopamine for Real-Time Determination of Carbohydrate–Lectin Interactions by QCM Biosensor

Polymers ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 1275 ◽  
Author(s):  
Kun Shang ◽  
Siyu Song ◽  
Yaping Cheng ◽  
Lili Guo ◽  
Yuxin Pei ◽  
...  
Keyword(s):  
Real Time ◽  
Con A ◽  
Quartz Crystals ◽  
Ulex Europaeus ◽  
Novel Approach ◽  
Ulex Europaeus Agglutinin I ◽  
Flow Through ◽  
Time Determination ◽  
Kinetics Of

A novel approach for preparing carbohydrate chips based on polydopamine (PDA) surface to study carbohydrate–lectin interactions by quartz crystal microbalance (QCM) biosensor instrument has been developed. The amino-carbohydrates were immobilized on PDA-coated quartz crystals via Schiff base reaction and/or Michael addition reaction. The resulting carbohydrate-chips were applied to QCM biosensor instrument with flow-through system for real-time detection of lectin–carbohydrate interactions. A series of plant lectins, including wheat germ agglutinin (WGA), concanavalin A (Con A), Ulex europaeus agglutinin I (UEA-I), soybean agglutinin (SBA), and peanut agglutinin (PNA), were evaluated for the binding to different kinds of carbohydrate chips. Clearly, the results show that the predicted lectin selectively binds to the carbohydrates, which demonstrates the applicability of the approach. Furthermore, the kinetics of the interactions between Con A and mannose, WGA and N-Acetylglucosamine were studied, respectively. This study provides an efficient approach to preparing carbohydrate chips based on PDA for the lectin–carbohydrate interactions study.

scholarly journals Analysis of human brain tissue derived from DBS surgery

2021 ◽  
Author(s):  
Salla M Kangas ◽  
Jaakko Teppo ◽  
Maija J Lahtinen ◽  
Anu Suoranta ◽  
Bishwa M Ghimire ◽  
...  
Keyword(s):  
Human Brain ◽  
Brain Tissue ◽  
Expression Data ◽  
Liquid Chromatography Mass Spectrometry ◽  
Human Brain Tissue ◽  
Specific Expression ◽  
Novel Approach ◽  
Neurosurgical Treatment ◽  
Disease Specific ◽  
Deep Brain

The implantation of deep brain stimulation (DBS) electrodes into the human brain is a neurosurgical treatment for, e.g., movement disorders. We describe a novel approach to collecting brain tissue from DBS surgery-guiding instruments for liquid chromatography-mass spectrometry and RNA sequencing analyses. Proteomics and transcriptomics showed that the approach is useful for obtaining disease-specific expression data. A comparison between our improved and the previous approaches and related datasets was performed.

CD47 Functionalization of Nanoparticles as a Poly(ethylene glycol) Alternative: A Novel Approach to Improve Drug Delivery

2021 ◽  
Vol 22 ◽  
Author(s):  
Noushin Rezaei Vandchali ◽  
Fatemeh Moadab ◽  
Eskandar Taghizadeh ◽  
Amir Tajbakhsh ◽  
Seyed Mohammad Gheibi-hayat
Keyword(s):  
Drug Delivery ◽  
Ethylene Glycol ◽  
Retention Time ◽  
Blood Circulation ◽  
Regulatory Protein ◽  
Reticuloendothelial System ◽  
Poly Ethylene Glycol ◽  
Drug Delivery Vehicles ◽  
Novel Approach ◽  
Poly Ethylene

Abstract: Bio-degradable nanoparticles (NPs) have several utilizations as the drug delivery vehicles due to their acceptable bio-availability, lower toxicity, potency for encapsulation and controlled release. Moreover, interaction of the NPs with the macrophages of reticuloendothelial system (RES) may decrease NPs efficacy for medical purposes. The surface of NPs is conventionally neutralized with the molecules such as poly(ethylene glycol) (PEG), as one of the most widely applied stealth polymers, in order to restrict the NPs clearance through the RES system. In fact, these molecules exhibit resistance to the RES clearance and proteins adsorption. It is unfortunate that modifying the PEG has some shortcomings like problems in the synthesis as well as correlation to the immune reaction. The CD47 receptor has been well known as a ‘don’t-eat-me’ molecule on the self-cells' surface. Therefore, the receptor will inhibit phagocytosis via binding to its ligand that is known as the signal regulatory protein α (SIRP-α). Moreover, the CD47 receptor, as one of the biomimetic substances, or its derivative peptides have been used recently on the surface of nanoparticles to inhibit phagocytosis and increase the NPs retention time in the blood circulation. Therefore, this review study examined the CD47 receptor and its role in the immune system as well as the use of the CD47 receptor in coating NPs to increase their retention time in the blood circulation.

A hollow fiber system for simple generation of human brain organoids

2017 ◽  
Vol 9 (9) ◽  
pp. 774-781 ◽  
Author(s):  
Yujuan Zhu ◽  
Li Wang ◽  
Fangchao Yin ◽  
Yue Yu ◽  
Yaqing Wang ◽  
...  
Keyword(s):  
Stem Cell ◽  
Human Brain ◽  
High Throughput ◽  
Hollow Fiber ◽  
Cell Biology ◽  
Hollow Fibers ◽  
Stem Cell Biology ◽  
Fiber System ◽  
Novel Approach ◽  
High Throughput Manner

Here we present a novel approach to engineer hiPSC-derived brain organoids within hollow fibers in a simple and high throughput manner by integrating biomaterials with stem cell biology.

Accurate retention time determination of co-eluting proteins in analytical chromatography by means of spectral data

2012 ◽  
Vol 110 (3) ◽  
pp. 683-693 ◽  
Author(s):  
Florian Dismer ◽  
Sigrid Hansen ◽  
Stefan Alexander Oelmeier ◽  
Jürgen Hubbuch
Keyword(s):  
Spectral Data ◽  
Retention Time ◽  
Time Determination

scholarly journals Glycan analysis of human neutrophil granules implicates a maturation-dependent glycosylation machinery

2020 ◽  
Vol 295 (36) ◽  
pp. 12648-12660 ◽  
Author(s):  
Vignesh Venkatakrishnan ◽  
Régis Dieckmann ◽  
Ian Loke ◽  
Harry C. Tjondro ◽  
Sayantani Chatterjee ◽  
...  
Keyword(s):  
Gradient Centrifugation ◽  
Effector Proteins ◽  
Protein Glycosylation ◽  
Sialyl Lewis X ◽  
Human Neutrophils ◽  
Immune Functions ◽  
Lewis X ◽  
Percoll Density Gradient Centrifugation ◽  
Percoll Density Gradient ◽  
And Function

Protein glycosylation is essential to trafficking and immune functions of human neutrophils. During granulopoiesis in the bone marrow, distinct neutrophil granules are successively formed. Distinct receptors and effector proteins, many of which are glycosylated, are targeted to each type of granule according to their time of expression, a process called “targeting by timing.” Therefore, these granules are time capsules reflecting different times of maturation that can be used to understand the glycosylation process during granulopoiesis. Herein, neutrophil subcellular granules were fractionated by Percoll density gradient centrifugation, and N- and O-glycans present in each compartment were analyzed by LC–MS. We found abundant paucimannosidic N-glycans and lack of O-glycans in the early-formed azurophil granules, whereas the later-formed specific and gelatinase granules and secretory vesicles contained complex N- and O-glycans with remarkably elongated N-acetyllactosamine repeats with Lewis epitopes. Immunoblotting and histochemical analysis confirmed the expression of Lewis X and sialyl-Lewis X in the intracellular granules and on the cell surface, respectively. Many glycans identified are unique to neutrophils, and their complexity increased progressively from azurophil granules to specific granules and then to gelatinase granules, suggesting temporal changes in the glycosylation machinery indicative of “glycosylation by timing” during granulopoiesis. In summary, this comprehensive neutrophil granule glycome map, the first of its kind, highlights novel granule-specific glycosylation features and is a crucial first step toward a better understanding of the mechanisms regulating protein glycosylation during neutrophil granulopoiesis and a more detailed understanding of neutrophil biology and function.

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