Preparation of Pyridylaminated O-Linked Sugar Chains from Glycoproteins Blotted on a Polyvinylidene Difluoride Membrane and Application to Human Granulocyte Colony-Stimulating Factor

Oh-eda, Masayoshi; Tominaga, Eri; Nabuchi, Yoshiaki; Matsuura, Tetsu; Ochi, Norimichi; Tamura, Masahiko; Hase, Sumihiro

doi:10.1006/abio.1996.0186

Cited by 16 publications

(12 citation statements)

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“…I neutrofili indotti con la somministrazione di lenograstim mostrano un'inalterata chemiotassi, al contrario dopo somministrazione di filgrastim i neutrofili mostrano una minor capacità chemiotattica (p = 0,008). Modificata da [15] midollo [11]. Filgrastim e il suo derivato pegilato sono molecole originate da cellule di Escherichia coli, diverse dalla molecola che normalmente è presente in natura.…”

Section: Efficaciaunclassified

Appropriate administration of Granulocyte-Colony Stimulating Factors

Rosti¹

2012

RHC

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Chemotherapy-induced febrile neutropenia is a potentially fatal complication of cancer treatment and is also the main reason of dose-reduction and/or delay of chemotherapy regimen. Prophylaxis with G-CSF is applicable to reduce the risk of chemotherapy-induced neutropenia. Two molecules of recombinant G-CSF are available for clinical use: lenograstim, identical to human native G-CSF, that is derived from mammalian cells and filgrastim, different to human native G-CSF, expressed in E. coli and commercialized in normal form and pegilated long-acting form. Neutrophil morphology and expected defense functions are modified by treatment with filgrastim, while they are not affected by lenograstim. These functionality differences observed in vitro are recently confirmed in a clinical trial that shows a lower incidence of febrile episodes with lenograstim compared to filgrastim in presence of G-CSF induced neutrophils. The long-term safety of lenograstim was supported by the results of a prospective, longer-term study involving almost 4,000 healthy donors. Another important question is the respect of timing of administration of G-CSF and chemotherapy. Absolutely in no case the plasma concentration of G-CSF is to be detected 48h before to 24h post chemotherapeutic drugs administration. In fact, this combination could result in an increased risk of mielotoxicity and a potential for an increase in sensitivity of rapidly dividing myeloid cells to cytotoxic-mutagenic chemotherapy potential. Lenograstim and filgrastim shows short half-life time, instead pegfilgrastim shows detectable concentrations for 16 days after a single administration. This is important to be considered, in particular in bi-weekly and tri-weekly adjuvant chemotherapy regimens. Anyway, the use of the lowest effective dose for the shortest adequate time of medications ensures the optimal balance among effectiveness, safety and costs of treatments, in a context that takes into account effectiveness and efficiency.Keywords Granulocyte-Colony Stimulating Factors; Lenograstim; Filgrastim; Pegfilgrastim; G-CSF; Timing; Appropriateness; Guidelines Appropriatezza d'uso dei fattori di crescita granulocitari nella profilassi e nel trattamento della neutropenia febbrile Introduzione Chemioterapia, neutropenia e G-CSFLa chemioterapia antiblastica, anche nell' era delle terapie target, resta il caposaldo del trattamento della maggioranza dei tumori solidi e delle neoplasie ematologiche. A fronte del beneficio clinico, alcuni effetti collaterali sono preminenti, in particolare la mielodepressione e, nell'ambito di questa, soprattutto la neutropenia. Il comparire di neutropenia durante il trattamento può portare una serie di ulteriori complicanze come la neutropenia febbrile e la riduzione della dose intensity, intesa come diminuzione della quantità di dose somministrata nell'unità di tempo. Da ciò deriva un possibile ridotto effetto dei farmaci sul controllo della malattia e quindi un peggiorato outcome [1]. Indubbiamente negli ultimi anni l'impiego in clinica...

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Section: Efficaciaunclassified

Appropriate administration of Granulocyte-Colony Stimulating Factors

Rosti¹

2012

RHC

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“…The low yield (8%) was attributed to inefficient electrotransfer, as 82% recovery of label was reported when the protein was directly applied to nylon or nitrocellulose membrane. The 0-linked oligosaccharides of human granulocyte-colony stimulating factor (G-CSF; 30 pg) were removed from a PVDF membrane by non-reductive 6-elimination followed by fluorescent derivatisation with pyridylamine [9]. The technique involved a 48 h reaction time with a yield of 55% and some loss of the reducing terminal N-acetyl galactosamine and terminal sialic acid.…”

Section: Introductionmentioning

confidence: 99%

Proteome analysis of glycoforms: A review of strategies for the microcharacterisation of glycoproteins separated by two‐dimensional polyacrylamide gel electrophoresis

Packer¹,

Pawlak²,

Kett³

et al. 1997

Electrophoresis

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Preparative two-dimensional polyacrylamide electrophoresis (2-D PAGE) is a method of separation which for the first time allows protein isoforms to be readily purified for subsequent analysis. The profile of the 2-D separation of the protein complement (proteome) of eukaryotic cells and tissues typically contains obvious 'trains' of spots which differ in pI and/or apparent molecular mass. These are usually isoforms of the same protein and result from post-translational modifications. There is growing evidence that alterations to the glycosylation and/or phosphorylation of a protein can be correlated with developmental and pathological changes; these changes can be visualised on the 2-D separation. It is not clear, however, how these modifications alter the structural properties of the protein and affect their migration in this mode of separation. Strategies need to be developed to obtain a more detailed understanding of the reason for the appearance of isoforms as discrete spots on 2-D PAGE. Standard proteins, fetuin and ovalbumin, were used to monitor the effect of the removal of glycans and phosphates on the migration of the glycoproteins in the 2-D system. The isoforms were not simply explained by the presence or absence of a single modification. To further investigate the reasons for the different migration of the isoforms it is necessary to characterise the modifications in more detail. Unlike protein analysis, until recently the available methodology for the analysis of the glycans attached to proteins has not been sensitive enough to allow analysis of single spots in gels or blots resulting from 2-D electrophoresis. In this paper we review current and future strategies for characterisation of protein modifications using single spots from 2-D gels.

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“…Another drawback of this procedure is that several noncarbohydrate substituents of the glycan chains may be destroyed. O-glycosidic linkages can be alternatively cleaved by the β-elimination reaction with or without concomitant reduction of the reducing sugar to an alditol [Geyer and Geyer, 1993;Oh-eda et al, 1996;Chai et al, 1997]. The great advantage of the latter method is that the reducing monosaccharide constituent is maintained intact, thus allowing subsequent labeling of the oligosaccharide (see below).…”

Section: Chemical Release Of Glycoprotein Glycansmentioning

confidence: 99%

Strategies for Glycoconjugate Analysis

Geyer

1998

Cells Tissues Organs

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Glycoconjugates such as glycoproteins and glycosphingolipids exhibit multiple functions in biological systems. A correlation of functional features with defined structural parameters, however, presupposes detailed information on the glycoconjugates’ carbohydrate moieties including the type of linkage of the oligosaccharide chain(s), monosaccharide composition as well as sequence, linkage positions and anomeric configurations of individual sugar monomers. Chemical and biochemical analyses are often impeded by the limited amounts of sample available and the vast structural heterogeneity of glycoconjugate glycans, thus requiring highly sensitive and efficient methods for detection, separation and structural elucidation of these carbohydrates. The aim of the article is to present suitable strategies for structural characterization of glycoconjugate glycans and to briefly review some of the techniques commonly used in this field.

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Preparation of Pyridylaminated O-Linked Sugar Chains from Glycoproteins Blotted on a Polyvinylidene Difluoride Membrane and Application to Human Granulocyte Colony-Stimulating Factor

Cited by 16 publications

References 4 publications

Appropriate administration of Granulocyte-Colony Stimulating Factors

Appropriate administration of Granulocyte-Colony Stimulating Factors

Proteome analysis of glycoforms: A review of strategies for the microcharacterisation of glycoproteins separated by two‐dimensional polyacrylamide gel electrophoresis

Strategies for Glycoconjugate Analysis

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