Engineered virus-like nanoparticles reverse heparin anticoagulation more consistently than protamine in plasma from heparin-treated patients

doi:10.1016/j.thromres.2011.03.021

Thrombosis Research

Volume 128, Issue 4, October 2011, Pages e9-e13

https://doi.org/10.1016/j.thromres.2011.03.021 Get rights and content

Abstract

Heparin is widely used for anticoagulation, often requiring the subsequent administration of a reversal agent. The only approved reversal agent for heparin is protamine sulfate, which induces well described adverse reactions in patients. Previously we reported a novel class of heparin antagonists based on the bacteriophage Qβ platform, displaying polyvalent cationic motifs which bind with high affinity to heparin. Here we report heparin reversal by the most effective of these virus-like particles (VLP) in samples from patients who were administered heparin during cardiac procedures or therapeutically for treatment of various thrombotic conditions. The VLP consistently reversed heparin in these samples, including those from patients that received high doses of heparin, with greater efficiency than a negative control VLP and with significantly less variability than protamine sulfate. These results provide the first step towards validation of heparin antagonist VLPs as viable alternatives to protamine.

Section snippets

Materials and methods

Virus-like particles (VLPs) derived from bacteriophage Qβ coat protein containing mutations of Thr18 to Arg (T18R) and of Lys16 to Met (K16M) were produced and isolated as previously described [15], [16]. Purified particles were stored in 0.1 M HEPES, pH 7.4.

Citrated plasma samples were collected from patients treated with intravenous unfractionated heparin (UFH) at Scripps Green Hospital after informed consent was obtained. Patients were identified and screened through the cardiac

Results

All of the therapeutic UFH samples were assayed for heparin reversal by the two VLPs using an aPTT assay. This assay was adapted for addition of VLP to the plasma samples, producing a reference clot time for pooled normal plasma of 46 seconds. Fig. 1A and B show heparin reversal by T18R and K16M respectively for one plasma sample from each patient as a representative subset of all the data collected. T18R significantly reduced the clotting time in the aPTT assay, indicating that it was

Discussion

The T18R VLP was found to reverse heparin anticoagulation at therapeutic levels significantly better than the negative control VLP, K16M (Fig. 1). Reversal was 69 ± 10% with a slight increase in reversal as UFH concentration increased. The 10% variability observed was remarkably low considering the expected differences in plasma from different patients. In a comparison of T18R to protamine sulfate with samples from patients treated with high doses of UFH during cardiac catheterization procedures,

Conflict of interest statement

We report no conflicts of interest.

Acknowledgements

This manuscript was supported by NIH/NCRR UL1 RR025774 (Pilot-Gale), The Stein Endowment Fund (Gale) and by NIH R01 CA112075 (Finn).

References (33)

S. Schulman et al.
Anticoagulants and their reversal
Transfus Med Rev
(2007)
S.E. Kimmel et al.
Adverse events after protamine administration in patients undergoing cardiopulmonary bypass: risks and predictors of under-reporting
J Clin Epidemiol
(1998)
D.J. Cushing et al.
Reversal of heparin-induced increases in aPTT in the rat by PM102, a novel heparin antagonist
Eur J Pharmacol
(2010)
Y. Byun et al.
Low molecular weight protamine: a potential nontoxic heparin antagonist
Thromb Res
(1999)
T.W. Wakefield et al.
A [+ 18RGD] protamine variant for nontoxic and effective reversal of conventional heparin and low-molecular-weight heparin anticoagulation
J Surg Res
(1996)
F. Ni Ainle et al.
Protamine sulfate down-regulates thrombin generation by inhibiting factor V activation
Blood
(2009)
C. Kearon et al.
Antithrombotic therapy for venous thromboembolic disease: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines (8th Edition)
Chest
(2008)
T.W. Wakefield et al.
Reversal of low-molecular-weight heparin anticoagulation by synthetic protamine analogues
J Surg Res
(1994)
P. Singh et al.
Bio-distribution, toxicity and pathology of cowpea mosaic virus nanoparticles in vivo
J Control Release
(2007)
A.C. Tissot et al.
Effect of immunisation against angiotensin II with CYT006-AngQb on ambulatory blood pressure: a double-blind, randomised, placebo-controlled phase IIa study
Lancet
(2008)

C. Peterson et al.

Improving heparin safety: A multidisciplinary invited conference

Hosp Pharm

(2008)

D.C. Classen et al.

Adverse drug events among hospitalized Medicare patients: epidemiology and national estimates from a new approach to surveillance

Jt Comm J Qual Patient Saf

(2010)

M.A. Crowther et al.

Heparin

R.N. Pike et al.

Control of the coagulation system by serpins. Getting by with a little help from glycosaminoglycans

FEBS J

(2005)

J.C. Horrow

Protamine: a review of its toxicity

Anesth Analg

(1985)

I.J. Welsby et al.

Hemodynamic changes after protamine administration: association with mortality after coronary artery bypass surgery

Anesthesiology

(2005)

Cited by (18)

Design and construction of bioinspired supramolecular self-assembled nanostructures
2022, Design, Principle and Application of Self-Assembled Nanobiomaterials in Biology and Medicine
Proteins are major structural and functional components of the cells. Supramolecular protein assemblies’ structural properties include their predefined size, shape, self-assembly, resistance, flexibility, versatility, symmetry, and polyvalence for functionalization or bioconjugation. The structural properties of supramolecular protein assemblies, such as their predefined nanometric size, make them versatile tools in nanotechnology and nanomedicine as biotemplates of scaffolds. Cytoskeleton proteins, protein cages, viruses, and virus-like particles are supramolecular assemblies composed of proteins that acquire a specific three-dimensional structure with highly symmetrical and monodisperse architectures with precise morphologies and ideal properties as multifunctional biotemplates. This chapter presents recent advances in the design and construction of bioinspired nanostructures with supramolecular self-assembly properties and discusses the properties and applications of the resulting integrative nanobiomaterials. These are particularly peptide-based, cytoskeleton proteins, and viral-derived nanostructures, including functionalization or bioconjugation with inorganic materials and nanoparticles. Furthermore, recent advances in de novo protein design are presented to develop novel and innovative protein-based materials that can solve different technical challenges, such as energy harvesting, energy conversion, self-assembly, drug delivery, and nanomedicine applications.
Engineered protein cages for selective heparin encapsulation
2021, Journal of Materials Chemistry B
Heparin antagonists: Alternatives to protamine
2021, Advances in Medicine and Biology. Volume 181
Targeting Biological Polyanions in Blood: Strategies toward the Design of Therapeutics
2020, Biomacromolecules
New long-term action insulin formulations obtained using polycations for heparin neutralization
2019, Bio-Algorithms and Med-Systems
Polyvalent Hybrid Virus-Like Nanoparticles with Displayed Heparin Antagonist Peptides
2018, Molecular Pharmaceutics

View all citing articles on Scopus

View full text

Regular ArticleEngineered virus-like nanoparticles reverse heparin anticoagulation more consistently than protamine in plasma from heparin-treated patients

Abstract

Section snippets

Materials and methods

Results

Discussion

Conflict of interest statement

Acknowledgements

Transfus Med Rev

J Clin Epidemiol

Eur J Pharmacol

Thromb Res

J Surg Res

Blood

Chest