Improved platelet storage using a sialidase inhibitor

A sialidase, platelet technology, applied in the preservation of human or animal body, medical preparations containing active ingredients, applications, etc., can solve problems such as platelet contamination

Inactive Publication Date: 2014-04-02
VELICO MEDICAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At current rates, one in 1,000 units to one in 2,000 units of platelets is sufficiently contaminated with bacteria to pose a significant risk to the recipient

Method used

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  • Improved platelet storage using a sialidase inhibitor
  • Improved platelet storage using a sialidase inhibitor
  • Improved platelet storage using a sialidase inhibitor

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0256] Example 1: Human platelets: Prolonged storage at room temperature and below room temperature results in loss of sialic acid and increased sialidase (neuraminidase) activity of human platelets loss

[0257] Platelets were stored at 4°C in the absence or presence of 1.2 mM nucleotide sugars and total sialic acid was quantified. Platelets were centrifuged, washed extensively, and resuspended in 140 mM NaCl, 3 mM KCl, 0.5 mM MgCl 2 , 5mM NaHCO 3 , 10mM glucose and 10mM HEPES, the pH was 7.4. Aliquots of resuspended platelets were lysed with RIPA buffer (Cell Signaling Technology) for protein quantification using the Pierce BCA protein assay kit or processed for quantification using QuantiChrom TM Sialic Acid Assay Kit Quantification of platelet sialic acid was performed according to the manufacturer's instructions (BioAssays Systems). The assay kit uses a modified Warren's method in which sialic acid is oxidized to formylpyruvate which reacts with thiobarbituric acid to...

example 2

[0262] Example 2: Mouse Platelets: Sialidase activity is increased during cold storage of mouse platelets and the sialidase inhibitor DANA increases the survival of mouse platelets in vivo.

[0263] Mouse platelet sialidase activity increased after 48 h of cold storage.

[0264] We have used the Amplex Red Neuraminidase (Sialidase) Assay Kit (Molecular probes, Eugene, OR, USA) to determine the presence of Sialidase surface activity in mouse platelets and after cooling and rewarming. Mouse platelets (2×10 9 ) were separated and suspended in the provided reaction buffer (0.5M Tris-HCl, pH7.2 and 1mM CaCl 2 )middle. Platelet-derived sialidase activity was measured over 2.5 h at room temperature. Figure 5 It was shown that sialidase activity was substantially increased after platelet storage at low temperature (4°C) compared to fresh room temperature platelets (RT). Crucially, sialidase activity was not derived from plasma because platelets were washed extensively prior to s...

example 3

[0270] Example 3: The role of sialylation / desialylation in defining the circulatory lifespan of platelets.

[0271] Human platelets produce Neu1 and Neu3 and release Neu1 into plasma.

[0272] The studies here suggest two novel mechanisms that contribute to the increased platelet clearance that occurs after storage. The first mechanism of platelet clearance is rapidly induced by cold storage in the absence of plasma when the GlcNAc residue on the N-linked glycan of GPIbα becomes exposed and is activated by the αMβ2 receptor on hepatic phagocytes The lectin domain recognition is mediated. The second clearance mechanism, induced by long-term storage of platelets in plasma at low temperature, starts slowly and occurs when GPIbα is desialylated and recognized by ASGP receptors on both hepatocytes and macrophages of the liver happens when. Recent data have revealed unexpected roles of endogenous sialidases and glycosyltransferases (GTs) in modulating the circulatory lifespan of ...

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Abstract

The present invention relates to a platelet additive solution (PAS) having an amount of one or more sialidase inhibitors and optionally one or more glycan-modifying agents; and one or more of PAS components that includes a salt, a citrate source, a carbon source, and any combination thereof. The present invention also relates to methods, compositions and kits for increasing the in vivo circulation time of isolated platelets by storing the platelets with one or more sialidase inhibitors. Additionally, the present invention relates to methods and compositions for reducing sialidase activity and inhibiting bacterial proliferation of one or more bacteria in a platelet product preparation from one or more donors.

Description

[0001] related application [0002] This application is a continuation-in-part of U.S. Application No. 13 / 474,473, filed May 17, 2012; a continuation-in-part of U.S. Application No. 13 / 474,627, filed May 17, 2012; and Continuation-in-Part of U.S. Application No. 13 / 474,679 filed and claiming the benefit of the following U.S. Provisional Application Nos.: U.S. Provisional Application No. 61 / 613,876 filed March 21, 2012; U.S. Provisional Application No. 61 / 613,876 filed March 21, 2012; Application No. 61 / 613,837; U.S. Provisional Application No. 61 / 503,984, filed July 1, 2011; and U.S. Provisional Application No. 61 / 487,077, filed May 17, 2011. [0003] The entire teaching content of the above application is hereby incorporated by reference. [0004] governmental support [0005] This invention was supported in whole or in part by grant number 3RO1HL089224-03S1 from the National Heart, Lung, and Blood Institute. This government agency has certain rights in this invention. Bac...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/00A61K35/14A01N1/02A61K35/19
CPCA61K35/19A01N1/0215A01N1/0226A61P7/04A61P43/00Y02A50/30A61K2300/00
Inventor 启勇彼得·刘K·霍夫麦斯特R·萨克斯坦
Owner VELICO MEDICAL
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