Canadian Blood Services Research and Development

Ottawa, Canada

Canadian Blood Services Research and Development

Ottawa, Canada
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Acker J.P.,University of Alberta | M.Croteau I.,Canadian Blood Services Research and Development | Yi Q.-L.,Canadian Blood Services Epidemiology
Clinica Chimica Acta | Year: 2012

Background: Percentage hemolysis in red cell concentrates (RCC) for transfusion is an indicator of RBC damage. As several factors need to be measured to determine hemolysis, and multiple assays are available for each, the choice of analytical methodology could critically influence results. Methods: Hemolysis was measured in 48 RCCs every 7. days during storage including expiry (42. days), with supernatant hemoglobin measured using the reference Drabkin's cyanmethemoglobin method and the Harboe spectrophotometric method, total hemoglobin measured using Drabkin's method and 3 automated analyzers (ADVIA 120, CELL-DYN 1700, Coulter AcT), and hematocrit measured using traditional centrifugation and automated analyzers. Results: The choice of method affects hemolysis measurement. Biases ranging from -. 0.01--. 0.03% were observed depending on the combination of methods used. Hematocrit measurement appeared to be a major determinant of bias, and the greatest bias was seen with the ADVIA 120 automated analyzer. Although results did not differ by a level thought to be of clinical significance, the choice of method will impact quality control pass/fail rates. Conclusion: Although guidelines exist in many jurisdictions regarding acceptable hemolysis levels in RCCs, these are silent regarding the methods to be used. The presence of bias highlights the need for standardization of methodology. © 2012 Elsevier B.V..

Sheffield W.P.,Canadian Blood Services Research and Development | Sheffield W.P.,McMaster University | Bhakta V.,Canadian Blood Services Research and Development | Talbot K.,Canadian Blood Services Research and Development | And 4 more authors.
Transfusion and Apheresis Science | Year: 2013

Background: Transfusable plasma is obtained by processing whole blood donations, by apheresis, or as solvent/detergent plasma (SD plasma), a pooled pathogen-reduced plasma product. The quality of plasma is typically assessed by testing the activities of multiple coagulation-related plasma proteins, due to a lack of clinical trial data linking plasma composition to clinical endpoints. We sought to update previous quality surveys of Canadian frozen plasma (FP; manufactured from single donor whole blood donation and frozen within 24. h of phlebotomy), to provide transfusionists with a more complete picture of its characteristics. Study design and methods: FP units (n=131) were tested for: the activity of factors V, VII, VIII, X, and XI, protein S (PS), α2-antiplasmin (AP), and fibrinogen; and the activated partial thromboplastin (APTT) and prothrombin (PT) times. Comparisons were made to: previous Canadian FP surveys; and to studies of single-donor plasma and SD plasma from other nations. Results: Mean FVIII, fibrinogen, or APTT values did not differ from the previous annual survey of Canadian FP; FV activity was increased and PT values decreased. FP produced with or without leukoreduction differed only in mean APTT. Canadian FP exhibited generally similar quality to that reported by other organizations in Europe and Asia for similarly manufactured single-donor plasma, but contained notably higher PS and AP (~four-fold) activities than did SD plasma. Conclusion: Our results indicate that Canadian FP is of similar quality to single-donor products produced in other jurisdictions. While it is of arguably superior in vitro quality to an SD plasma product recently licensed in Canada, these differences are highly unlikely to have clinical significance for most indications for plasma transfusion. © 2013 Elsevier Ltd.

Culibrk B.,University of British Columbia | Stone E.,University of British Columbia | Levin E.,University of British Columbia | Weiss S.,Canadian Blood Services Research and Development | And 2 more authors.
Vox Sanguinis | Year: 2012

Background and Objectives There is no automated, accurate assay for the enumeration of residual red blood cells (rRBCs) in non-RBC components for transfusion, despite the potential risk of allo-immunization when mismatched components are transfused. Materials and Methods The automated ADVIA 120 cerebrospinal fluid (CSF) assay, which is approved to count RBCs and WBCs in CSF samples, was optimized and tested to measure rRBC in platelet concentrate (PC) and plasma components. Results Sample dilution, incubation time and reagent volume were optimized for use with non-RBC blood products. The assay was linear (R2=0·99), even at low rRBCs counts. Intra- and inter-assay variation gave coefficients of variance (CV) between 2·2 and 9·4% and 2·6 and 14·9%, respectively, depending on rRBC levels. Good correlation (r=0·995) was found between the automated assay and manual counting, which is considered the gold standard. Using the automated assay, the range of rRBCs (count/unit) in buffy-coat platelet concentrate (PCs) was 27-5505×106 and in apheresis PCs was 1-361×106. Conclusion The ADVIA CSF assay is a sensitive, precise and accurate means to assess rRBC counts in non-RBC components. © 2012 International Society of Blood Transfusion.

Sheffield W.P.,Canadian Blood Services Research and Development | Sheffield W.P.,McMaster University | Bhakta V.,Canadian Blood Services Research and Development | Trigatti B.,McMaster University | Jenkins C.,Canadian Blood Services Research and Development
Vox Sanguinis | Year: 2011

Background and Objective Cryoprecipitate prepared from two whole blood donations from the same donor contained insoluble orange particulate material (OPM). We sought to identify the OPM. Materials and Methods OPM was recovered from the blood product by centrifugation, dissolved in sodium dodecyl sulphate (SDS) and analysed by SDS-polyacrylamide gel electrophoresis and immunoblotting. Results Solubilized OPM was enriched in apolipoproteins B and E, but not apolipoprotein A1, immunoglobulin G or albumin, suggesting lipoprotein enrichment in OPM. Subsequent clinical laboratory blood tests confirmed low-density lipoprotein hyperlipidaemia with normal triglyceride levels. Further, cryoprecipitate production from this donor was prevented by implementation of national predominantly male plasma policies. Conclusion Cryoprecipitate produced from hyperlipidaemic donors may contain insoluble particles that render it inappropriate for transfusion. © 2010 The Author(s). Vox Sanguinis © 2010 International Society of Blood Transfusion.

Sheffield W.P.,McMaster University | McCurdy T.R.,McMaster University | Bhakta V.,Canadian Blood Services Research and Development | Eltringham-Smith L.J.,McMaster University | And 2 more authors.
Journal of Biomedicine and Biotechnology | Year: 2011

Alpha-1 acid glycoprotein (AGP) is a highly glycosylated, negatively charged plasma protein suggested to have anti-inflammatory and/or immunomodulatory activities. Purification of AGP could be simplified if methods that exploit its high solubility under chemically harsh conditions could be demonstrated to leave the protein in its native conformation. Procedures involving exposure of AGP to hot phenol or sulphosalicylic acid (SSA) were compared to solely chromatographic methods. Hot phenol-purified AGP was more rapidly cleared from mice in vivo following intravenous injection than chromatographically purified AGP. In contrast, SSA-purified AGP demonstrated an identical in vivo clearance profile and circular dichroism spectrum to chromatographically purified AGP. Similarly, no differences in susceptibility to enzymatic deglycosylation or reactivity with Sambucus nigra lectin were detected between AGP purified via the two methods. Incorporation of the SSA step in the purification scheme for AGP eliminated the need for a large (4mL resin/mL of plasma) initial chromatographic step and simplified its purification without causing any detectable distortion in the conformation of the protein. Confirmation that this procedure is nondenaturing will simplify AGP purification and investigation of its possible biological roles in laboratory animals. Copyright 2011 Teresa R. McCurdy et al.

Mccurdy T.R.,McMaster University | Patrick A.L.,McMaster University | Eltringham-Smith L.J.,McMaster University | Bhakta V.,Canadian Blood Services Research and Development | And 2 more authors.
Microcirculation | Year: 2014

Objective: To characterize the effect of systemically administered AGP on early leukocyte recruitment in the livers of endotoxemic or septic mice and to determine whether this is influenced by LPS sequestration. Methods: Endotoxemia was induced in C57Bl/6 mice via intraperitoneal injection of LPS. Sepsis was induced in mice by cecal ligation and perforation. AGP (165 mg/kg) or saline (20 mL/kg) or HAS (200 mg/kg) was administered immediately after surgery or LPS injection and the hepatic microcirculation was examined by intravital microscopy at four hour. Results: Leukocyte adhesion in the PSV was reduced by treatment with AGP in mice subjected to either LPS or CLP protocols compared to either saline or HAS treatment. AGP-treated mice also had significantly higher sinusoidal flow in both models. Pre-incubation of LPS with AGP reduced the ability of LPS to recruit leukocytes to the liver microcirculation. Conclusions: AGP was more effective in limiting hepatic inflammation and maintaining perfusion than saline or HAS, in both endotoxemic and septic mice. AGP sequestration of LPS may contribute to its anti-inflammatory effects. © 2013 John Wiley & Sons Ltd.

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