Further characterization of the binding of heparin to granulocyte colony‐stimulating factor: Importance of sulfate groups

Liang, Aiye; Liu, Xiaojun; Du, Yifeng; Wang, Keyi; Lin, Bingcheng

doi:10.1002/elps.200700480

Cited by 6 publications

(3 citation statements)

References 43 publications

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“…Thereby GAG-bind proteins are protected against proteases and interactions with their receptors are facilitated [ 9 , 32 , 33 ]. For example, fine characterization of the binding of heparin to G-CSF demonstrated the importance of sulfate groups since 2,3- O -sulfate groups are more important than N -sulfate groups in heparin–G-CSF interaction [ 34 ]. Heparin-induced leukocytosis requires glucosamine 6- O -sulfation and is caused by blockade of L-selectin-mediated, P-selectin-mediated, and CXCL12-mediated leukocyte trafficking [ 35 ].…”

Section: Discussionmentioning

confidence: 99%

In vitro and in vivo evaluation of cord blood hematopoietic stem and progenitor cells amplified with glycosaminoglycan mimetic

et al. 2016

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BackgroundExpansion protocols aim at both increasing the number of umbilical cord blood (UCB) hematopoietic stem cells and progenitor cells (HSPCs) and reducing the period of neutropenia in UCB HSPC graft. Because glycosaminoglycans (GAGs) are known to be important components of the hematopoietic niche and to modulate growth factor effects, we explored the use of GAG mimetic OTR4131 to potentiate HSPC’s in vitro expansion and in vivo engraftment.MethodsUCB CD34+ cells were expanded with serum-free medium, SCF, TPO, FLT3-lig and G-CSF during 12 days in the absence or the presence of increasing OTR4131 concentrations (0-100 μg/mL). Proliferation ratio, cell viability and phenotype, functional assays, migration capacity and NOD-scid/γc-/- mice engraftment were assessed after expansion.ResultsAt Day 12, ratios of cell expansion were not significantly increased by OTR4131 treatment. Better total nucleated cell viability was observed with the use of 1 μg/mL GAG mimetic compared to control (89.6 % ± 3.7 % and 79.9 % ± 3.3 %, respectively). Phenotype analysis showed a decrease of monocyte lineage in the presence of OTR4131 and HSPC migration capacity was diminished when GAG mimetic was used at 10 μg/mL (10.9 % ± 4.1 % vs. 52.9 % ± 17.9 % for control). HSPC clonogenic capacities were similar whatever the culture conditions. Finally, in vivo experiments revealed that mice successfully engrafted in all conditions, even if some differences were observed during the first month. Three months after graft, bone marrow chimerism and blood subpopulations were similar in both groups.ConclusionsUCB HSPCs ex-vivo expansion in the presence of OTR4131 is a safe approach that did not modify cell function and engraftment capacities. In our experimental conditions, the use of a GAG mimetic did not, however, allow increasing cell expansion or optimizing their in vivo engraftment.

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Section: Discussionmentioning

confidence: 99%

In vitro and in vivo evaluation of cord blood hematopoietic stem and progenitor cells amplified with glycosaminoglycan mimetic

et al. 2016

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“…CE-coupled with laser-induced fluorescence (LIF) detection has allowed for detection and quantitation of cytokines, chemokines, neuropeptides, and other biomarkers found in clinical samples (1–3). Immunoaffinity capillary electrophoresis (ICE) is a customized technique that tailors the capillary to capture the analytes of interest.…”

Section: Introductionmentioning

confidence: 99%

Cytokine Analysis by Immunoaffinity Capillary Electrophoresis

Mendonca

Kalish

2012

Methods in Molecular Biology

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Immunoaffinity capillary electrophoresis (ICE) is a powerful tool used to detect and quantify target proteins of interest in complex biological fluids. The target analyte is captured and bound to antibodies immobilized onto the wall of a capillary, labeled in situ with a fluorescent dye, eluted and detected online using laser-induced fluorescence following electrophoretic separation. Here, we illustrate how to construct an immunoaffinity capillary and utilize it to run ICE in order to capture and quantify target cytokines and chemokines from a clinical sample.

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“…Further, a number of protocols relying on HPLC 11, 12, NMR 13–15, CE 16, 17, and a combination of techniques 18 have been developed for detecting UFH contaminants. CE in particular has proven itself in the analysis of heparin disaccharides and oligosaccharides 19, and in the characterization of interaction of different proteins with heparin‐like highly charged molecules 20–23. CE‐based intact GAG analysis typically results in a broad, featureless profile that spans several minutes 24.…”

Section: Introductionmentioning

confidence: 99%

Linear polyalkylamines as fingerprinting agents in capillary electrophoresis of low‐molecular‐weight heparins and glycosaminoglycans

King¹,

Desai²

2011

Electrophoresis

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Glycosaminoglycan (GAG) analysis represents a challenging frontier despite the advent of many high resolution technologies because of their unparalleled structural complexity. We previously developed a resolving agent aided capillary electrophoretic approach for fingerprinting low molecular weight heparins (LMWHs) to profile their microscopic differences and assess batch-to-batch variability. In this work, we study the application of this approach for fingerprinting other GAGs and analyze the basis for the fingerprints observed in CE. Whereas the resolving agents, linear polyalkylamines, could resolve the broad featureless electropherogram of LMWH into a large number of distinct, highly reproducible peaks, longer GAGs such as chondroitin sulfate, dermatan sulfate and heparin responded in a highly individualistic manner. Full-length heparin interacted with linear polyalkylamines very strongly followed by dermatan sulfate, while chondroitin sulfate remained essentially unaffected. Oversulfated chondroitin sulfate could be easily identified from full-length heparin. Scatchard analysis of the binding profile of enoxaparin with three linear polyalkylamines displayed a biphasic binding profile suggesting two distinctly different types of interactions. Some LMWH chains were found to interact with linear polyalkylamines with affinities as high as 10 nM, while others displayed nearly 5000-fold weaker affinities. These observations provide fundamental insight into the basis for fingerprinting of LMWHs by linear polyalkylamine-based resolving agents, which could be utilized in the design of advanced resolving agents for compositional profiling, direct sequencing and chemoinformatics studies.

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Further characterization of the binding of heparin to granulocyte colony‐stimulating factor: Importance of sulfate groups

Cited by 6 publications

References 43 publications

In vitro and in vivo evaluation of cord blood hematopoietic stem and progenitor cells amplified with glycosaminoglycan mimetic

In vitro and in vivo evaluation of cord blood hematopoietic stem and progenitor cells amplified with glycosaminoglycan mimetic

Cytokine Analysis by Immunoaffinity Capillary Electrophoresis

Linear polyalkylamines as fingerprinting agents in capillary electrophoresis of low‐molecular‐weight heparins and glycosaminoglycans

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