Use of a Design of Experiments (DoE) Approach to Optimize Large-Scale Freeze-Thaw Process of Biologics

2021 ◽  
Vol 22 (4) ◽  
Author(s):  
Bruna Minatovicz ◽  
Robin Bogner ◽  
Bodhisattwa Chaudhuri
Keyword(s):  
Design Of Experiments ◽  
Large Scale ◽  
Freeze Thaw

scholarly journals Effects of Serum Stored at Various Temperature and FreezeThaw Cycles on Selected Biochemical Parameters

2016 ◽  
Vol 15 (1) ◽  
Author(s):  
Nor Zamzila Abdullah ◽  
Norlelawati A. Talib ◽  
Nor Sidah Ku Zaifah ◽  
Abdul Hadi Mohamed ◽  
Naznin Muhammad ◽  
...  
Keyword(s):  
Biochemical Parameters ◽  
Large Scale ◽  
Storage Temperature ◽  
Density Lipoprotein ◽  
Storage Conditions ◽  
Freeze Thaw ◽  
Thaw Cycle ◽  
Freeze Thaw Cycle ◽  
Extended Duration ◽  
Positive Correlations

Introduction: Preanalytical variation as a result of sample storage conditions are common issues in a large scale study since the sample has to be kept for an extended duration at ultra low temperature (-70ºC) before analysis. However, ultra low freezers may not be available in some laboratories. Sometimes, the laboratory test may need to be repeated on the previously frozen and thawed samples. The aim of this research was to study the effect of storage temperature and freeze-thaw cycles on some biochemical parameters in our laboratory. Methods: An experimental study was carried out on 50 volunteers. Serum were aliquoted and stored at 4ºC, -30ºC and -70ºC. The serum was analyzed for total cholesterol (TC), triglycerides (TG), high density-lipoprotein (HDL-C) and glucose after 3 months of storage. The effects of freeze-thaw cycles were also recorded. Results: No significant differences (p>0.05) were seen in the level of the biochemical parameters between the samples stored at -70ºC and at -30ºC. The levels of selected parameters were higher after second freeze-thaw cycles as compared to the first thaw. All parameters showed significant positive correlations between the samples stored at -70ºC and the samples stored at -30ºC and 4ºC as well as between the first and second freeze-thaw cycles in samples kept at both -70ºC and -30ºC. Conclusions: Our finding suggested that the level of TC, TG and glucose in sample stored for 3 months at -70ºC are comparable with the samples stored at -30ºC. Only one freeze-thaw cycle is acceptable.

scholarly journals Influence of viral transport media and freeze-thaw cycling on the sensitivity of qRT-PCR detection of SARS-CoV-2 nucleic acids

2021 ◽  
Author(s):  
Cian Holohan ◽  
Sophia Hanrahan ◽  
Nathan Feely ◽  
Peng Li ◽  
John O'Connell ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Large Scale ◽  
Pcr Detection ◽  
Molecular Diagnostic ◽  
Pcr Analysis ◽  
Analytical Sensitivity ◽  
Freeze Thaw ◽  
Qrt Pcr ◽  
Viral Transport ◽  
The Stability

The events of the last year have highlighted the complexity of implementing large-scale molecular diagnostic testing for novel pathogens. The purpose of this study was to determine the chemical influences of sample collection media and storage on the stability and detection of viral nucleic acids by qRT-PCR. We studied the mechanism(s) through which viral transport media (VTM) and number of freeze-thaw cycles influenced the analytical sensitivity of qRT-PCR detection of SARS-CoV-2. Our goal is to reinforce testing capabilities and identify weaknesses that could arise in resource-limited environments that do not have well-controlled cold chains. The sensitivity of qRT-PCR analysis was studied in four VTM for synthetic single-stranded RNA (ssRNA) and double-stranded DNA (dsDNA) simulants of the SARS-CoV-2 genome. The sensitivity and reproducibility of qRT-PCR for the synthetic ssRNA and dsDNA were found to be highly sensitive to VTM with the best results observed for ssRNA in HBSS and PBS-G. Surprisingly, the presence of epithelial cellular material with the ssRNA increased the sensitivity of the qRT-PCR assay. Repeated freeze-thaw cycling decreased the sensitivity of the qRT-PCR with two noted exceptions. The choice of VTM is critically important to defining the sensitivity of COVID-19 molecular diagnostics assays and this study suggests they can impact upon the stability of the SARS-CoV-2 viral genome. This becomes increasingly important if the virus structure is destabilised before analysis, which can occur due to poor storage conditions. This study suggests that COVID-19 testing performed with glycerol-containing PBS will produce a high level of stability and sensitivity. These results are in agreement with clinical studies reported for patient-derived samples.

Simulation-driven design of experiments examining the large-scale, explosive dispersal of particles

Shock Waves ◽  
2019 ◽  
Vol 30 (4) ◽  
pp. 325-347 ◽  
Author(s):  
K. T. Hughes ◽  
S. Balachandar ◽  
A. Diggs ◽  
R. Haftka ◽  
N. H. Kim ◽  
...  
Keyword(s):  
Design Of Experiments ◽  
Large Scale ◽  
Explosive Dispersal

Influence on Pitting Corrosion Behavior of Welded AISI30400 Plate in a Freeze-Thaw Environment

2014 ◽  
Vol 590 ◽  
pp. 307-311
Author(s):  
Tsuyoshi Takahashi ◽  
Yoshitaka Iwabuchi ◽  
Kazunori Ishitsuka ◽  
Keita Murakami
Keyword(s):  
Residual Stress ◽  
Stainless Steel ◽  
Austenitic Stainless Steel ◽  
Pitting Corrosion ◽  
Large Scale ◽  
Chloride Ions ◽  
Temperature Cycling ◽  
Local Concentration ◽  
X Rays ◽  
Freeze Thaw

Welded structures which set up in outdoor in snow-and cold-prone areas are exposed to severe environment, in which liquids containing Cl-chloride ions from the sea breeze and snow melting agents undergo freeze-thaw cycles during the winter. Cl-solutions are known to produce pitting corrosion in austenitic stainless steel. However, the relationship between the freeze-thaw environment and pitting corrosion occurred in welded austenitic stainless steel has not yet been clarified. In the present study, the effect of the freeze-thaw environment on pitting corrosion in welded austenitic stainless steel submersed in Cl-solutions is investigated. AISI 304 (X5CrNi18-10 in ISO/TS 15510, SUS304 in JIS G 4305) samples with bead-on-plate welding are used as specimens. The thickness of the specimen is 4 mm. The specimens are fully immersed in FeCl3 solution. The freeze-thaw environment is created by temperature cycling from 20°C to-20°C. In the meanwhile, residual stresses due to the welding process in stainless steel structures can significantly affect behavior of pitting corrosion as same as influence of sensitization. Authors carried out the welding simulation as the purpose of predicting the welding-heat history and residual stress near the welding bead. As a result, large-scale pitting corrosion occurs primarily in areas of high tensile residual stress and sensitive metallographic structures near weld beads in the freeze-thaw environment. Accelerated pitting corrosion is confirmed to have occurred as a result of concentration cell corrosion due to the local concentration of Cl-chlorine ion around a weld bead during freezing and thawing of the solution. Furthermore, based on your study it became clear that the center position of large-scale pitting corrosion is in agreement with the simulation result rather than the stress peak position by X rays measurement.

scholarly journals The Effects of a Single Freeze-Thaw Cycle on Concentrations of Nutritional, Noncommunicable Disease, and Inflammatory Biomarkers in Serum Samples

2021 ◽  
Author(s):  
Ransi Ann Abraham ◽  
Garima Rana ◽  
Praween K. Agrawal ◽  
Robert Johnston ◽  
Avina Sarna ◽  
...  
Keyword(s):  
Large Scale ◽  
Low Density Lipoprotein ◽  
Density Lipoprotein ◽  
Noncommunicable Disease ◽  
Noncommunicable Diseases ◽  
Serum Samples ◽  
Freeze Thaw ◽  
Thaw Cycle ◽  
Freeze Thaw Cycle ◽  
The Stability

Abstract Background The stability of biological samples is vital for reliable measurements of biomarkers in large-scale survey settings, which may be affected by freeze-thaw procedures. We examined the effect of a single freeze-thaw cycle on 13 nutritional, noncommunicable diseases (NCD), and inflammatory bioanalytes in serum samples. Method Blood samples were collected from 70 subjects centrifuged after 30 minutes and aliquoted immediately. After a baseline analysis of the analytes, the samples were stored at − 70°C for 1 month and reanalyzed for all the parameters. Mean percentage differences between baseline (fresh blood) and freeze-thaw concentrations were calculated using paired sample t-tests and evaluated according to total allowable error (TEa) limits (desirable bias). Results Freeze-thaw concentrations differed significantly (p < 0.05) from baseline concentrations for soluble transferrin receptor (sTfR) (− 5.49%), vitamin D (− 12.51%), vitamin B12 (− 3.74%), plasma glucose (1.93%), C-reactive protein (CRP) (3.45%), high-density lipoprotein (HDL) (7.98%), and cholesterol (9.76%), but they were within respective TEa limits. Low-density lipoprotein (LDL) (− 0.67%), creatinine (0.94%), albumin (0.87%), total protein (1.00%), ferritin (− 0.58%), and triglycerides (TAG) (2.82%) concentrations remained stable following the freeze-thaw cycle. In conclusion, single freeze-thaw cycle of the biomarkers in serum/plasma samples after storage at − 70°C for 1 month had minimal effect on stability of the studied analytes, and the changes in concentration were within acceptable limit for all analytes.

Report on Session 2a

1991 ◽  
Vol 7 (1) ◽  
pp. 335-337 ◽  
Author(s):  
K. D. Privett
Keyword(s):  
Natural Condition ◽  
Large Scale ◽  
Engineering Properties ◽  
Frost Heave ◽  
List Type ◽  
Freeze Thaw ◽  
Materials Engineering ◽  
Loess Deposition ◽  
Geomorphological Processes ◽  
Lake Deposits

IntroductionThe papers in this session of the conference concentrate on some of the processes described by the Theme Lecturer, Professor Hutchinson, and the engineering properties of the materials. Engineering implications are referred to in the papers but are included in this session because the process/properties element is stressed. This report is intended to provide a brief introduction to the session papers and all references are to papers in this conference.The term “periglacial” is sometimes restricted to geomorphological processes where freeze-thaw is the dominant action but the definition generally is widened to include all those processes, and their landforms, taking place in cold climated outside the margins of an ice sheet and so encompasses everything that is not directly glacial. This conference has used this wider definition and thus deposits such as laminated glacial lake sediments (Bell and Coultard) are included in this session. Table 1 illustrated the range of features that could be considered, many of which are discussed to varying degress in the papers.TABLE 1: List of periglacial features/processes.LARGE-SCALE LANDSLIPSSOLIFLUCTION, MUDFLOWS, SHEARSASYMMETRIC VALLEYSFROST CREEPCAMBERS, VALLEY BULGESICE WEDGESPOLYGONSFROST HEAVE, INVOLUTIONSFROST MOUNDS, PINGOSFROST SHATTERINGLOESS DEPOSITIONLAMINATED LAKE DEPOSITSCHEMICAL OF CaCO3SLOPE PROCESSESThe various processes involved in the formation and modification of slopes, either in their natural condition, or if modified by some engineering works. Is is not surprising then, that the majority of papers submitted to this session concern periglacial slope processes.

scholarly journals SURFACE DEFORMATION MONITORING IN PERMAFROST REGIONS OF TIBETAN PLATEAU BASED ON ALOS PALSAR DATA

2017 ◽  
Vol XLII-2/W7 ◽  
pp. 1509-1512
Author(s):  
L. M. Chen ◽  
G. Qiao ◽  
P. Lu
Keyword(s):  
Structural Damage ◽  
Large Scale ◽  
Surface Deformation ◽  
Deformation Monitoring ◽  
Frozen Soil ◽  
Tibet Plateau ◽  
Temperature Deformation ◽  
Deformation Pattern ◽  
Permafrost Region ◽  
Freeze Thaw

The permafrost region of Qinghai-Tibet Plateau is widely distributed with the freeze/thaw processes that cause surface structural damage. The differential interferometry synthetic aperture radar (DInSAR) can detect large scale surface deformation with high precision, thus can be used to monitor the freeze/thaw processes of frozen soil area. In this paper, the surface deformation pattern of Qinghai-Tibet railway was analyzed by using the PALSAR 1.0 raw data of the ALOS satellite (L band) and 90m resolution SRTM DEM data, with the help of two-pass DInSAR method in GAMMA software, and the differential interferograms and deformation maps were obtained accordingly. Besides, the influence of temperature, topography and other factors on deformation of frozen soil were also studied. The following conclusions were obtained: there is a negative correlation between deformation and temperature, and there is a delay between the deformation change and that of temperature; deformation and elevation are positively correlated; the permafrost deformation is also affected by solar radiation that could form variable amplitude variation.

scholarly journals Influence of viral transport media and freeze-thaw cycling on the sensitivity of qRT-PCR detection of SARS-CoV-2 nucleic acids

Nanoscale ◽  
2021 ◽  
Author(s):  
Cian Holohan ◽  
Sophia Hanrahan ◽  
Nathan Feely ◽  
Peng Li ◽  
John O'Connell ◽  
...  
Keyword(s):  
Infectious Diseases ◽  
Nucleic Acids ◽  
Large Scale ◽  
Diagnostic Testing ◽  
Pcr Detection ◽  
Molecular Diagnostic ◽  
Freeze Thaw ◽  
Qrt Pcr ◽  
Viral Transport ◽  
Molecular Diagnostic Testing

Objective The events of the last year have highlighted the complexity of implementing molecular diagnostic testing for infectious diseases at a large scale. The purpose of this study was to...

scholarly journals Stability of Soluble Adhesion Molecules, Selectins, and C-Reactive Protein at Various Temperatures: Implications for Epidemiological and Large-Scale Clinical Studies

2007 ◽  
Vol 53 (10) ◽  
pp. 1858-1860 ◽  
Author(s):  
Janine Hartweg ◽  
Michael Gunter ◽  
Rafael Perera ◽  
Andrew Farmer ◽  
Carole Cull ◽  
...  
Keyword(s):  
Adhesion Molecules ◽  
Whole Blood ◽  
Large Scale ◽  
Venous Blood ◽  
Storage Conditions ◽  
C Reactive Protein ◽  
Soluble Adhesion Molecules ◽  
Freeze Thaw ◽  
Reactive Protein ◽  
The Impact

Abstract Background: We assessed the impact of sample storage conditions on soluble vascular cell adhesion molecules (sVCAM), soluble intracellular adhesion molecules (sICAM-1), soluble (s)E-selectin, C-reactive protein (CRP), and sP-selectin. Methods: Markers were measured by ELISA in venous blood from 10 healthy volunteers on aliquots stored as plasma or whole blood at 4, 21, or 30 °C for 1–5 days and after 1–5 freeze-thaw cycles. We compared results on these samples to results for samples processed immediately and stored at −80 °C. Statistical models assessed time-related effects and effects of postprocessing conditions. Results: Using an upper limit of 10% variation from baseline with P &gt;0.05, we found that stability duration in plasma was 5 days for sVCAM-1 and sICAM-1 and at least 2 days for sE-selectin at 4, 21, and 30 °C and 5 days for CRP at 4 and 21 °C and 1 day at 30 °C. Stability duration in whole blood was 5 days for sVCAM-1 and sICAM-1 and at least 2 days for sE-selectin at 4, 21, and 30 °C and 5 days for CRP at 4 and 21 °C and 2 days at 30 °C. sP-selectin was not stable in plasma or whole blood. sICAM-1, sVCAM-1, CRP, and sE-selectin were stable after 5 freeze-thaw cycles. Conclusions: sVCAM-1, sICAM-1, and CRP are stable in plasma or whole blood at 4 and 21 °C for at least 3 days and sE-selectin for 2 days. sP-selectin is not stable and therefore requires immediate assay.

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