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To evaluate the effects of ketamine on assay performance (versus in vivo physiologic effects), three serum samples, each taken from a different cooperating cynomolgus macaque were measured neat (unspiked), and spiked with ketamine. Ketamine was diluted to 0.02 mg/mL with kit-provided sample matrix, and added to serum samples to give a final concentration of 2000 ng/mL, the reported peak concentration of ketamine in human plasma during sedation 26. Samples were tested in duplicate on a single assay plate. Data analysis Johnson-Davis KL, Farnsworth C, Law C, Parker R. Method validation for a multi-element panel in serum by inductively coupled plasma mass spectrometry (ICP-MS). Clin Biochem. 2020;82:90–8. Chen J-L, Guo Y-L, Tsai P-J, Su L-F. Use of inhalable Cr+ 6 exposures to characterize urinary chromium concentrations in plating industry workers. J Occup Health. 2002;44(1):46–52.
serum and plasma matrices on multiplex Effects of serum and plasma matrices on multiplex
Gibbs, R. A. et al. Evolutionary and biomedical insights from the rhesus macaque genome. Science 316(5822), 222–234. https://doi.org/10.1126/science.1139247 (2007) (Epub 2007/04/14). We are very pleased to see the enthusiasm with which the communities of Bougouni received the first doses of this new candidate malaria vaccine. We are hopeful that the encouraging Phase 2 results will be confirmed in this larger Phase 3 trial'. M. Bost, S. Houdart, M. Oberli, E. Kalonji, J.-F. Huneau and I. Margaritis, J. Trace Elem. Med. Biol., 2016, 35, 107–115 CrossRef CAS PubMed. Multiplexing cytokine measurements is an important research technique that improves efficiency, preserves biological samples allowing for reduction in number of animal subjects needed, and gives a wealth of complex information about the clinical state of a subject. This study outlines the limitations and challenges involved in detection and interpretation of multiple cytokines in healthy NHP serum, as well as the variability in cytokine expression due to species, sex, and sedation. Of the 23 cytokines validated in healthy NHP serum for the Millipore Milliplex assay, 11 met all acceptance criteria (TGFα, IFNγ, IL-2, IL-15, IL-13, IL-5, IL-6, IL-8, MCP-1, IL-12/23 (p40) and VEGF). Five were marginally accepted as they didn’t meet acceptance criteria for either recovery or linearity (G-CSF, IL-10, sCD40L, IL-4 and MIP-1α). Seven were rejected as they didn’t meet acceptance criteria for both recovery and linearity (GM-CSF, IL-17A, IL-1ra, IL-1β, TNFα, MIP-1β and IL-18), indicating that these analytes may not be reliably detected using this method in healthy NHP serum. First vaccinations have now begun in Mali in a phase III trial of a malaria vaccine developed at the University of Oxford. Known as R21/Matrix-M, it recently showed efficacy of 77% over 12 months in a phase IIb trial, and it is hoped that this phase III trial will help to lead to licensure of this malaria vaccine by 2023.
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Doe, R. P., Vennes, J. A. & Flink, E. B. Diurnal variation of 17-hydroxycorticosteroids, sodium, potassium, magnesium and creatinine in normal subjects and in cases of treated adrenal insufficiency and Cushing’s syndrome. J. Clin. Endocrinol. Metab. 20, 253–265. https://doi.org/10.1210/jcem-20-2-253 (1960). Of the 23 cytokines that underwent validation in healthy NHP serum, 11 met all acceptance criteria, 5 were marginally accepted, and 7 were rejected. These results demonstrate that the Millipore Milliplex multiplex assay can be reliably used to detect each of the 11 accepted cytokines (TGFα, IFNγ, IL-2, IL-15, IL-13, IL-5, IL-6, IL-8, MCP-1, IL-12/23 (p40) and VEGF) at endogenous levels in serum from healthy NHPs. The marginal cytokines meeting most of the validation criteria (G-CSF, IL-10, sCD40L, IL-4 and MIP-1α) require consideration of method specific limitations prior to interpretation of absolute measurements. The 7 cytokines in the panel that did not meet our pre-established acceptance criteria (GM-CSF, IL-17A, IL-1ra, IL-1β, TNFα, MIP-1β and IL-18) may not be reliably detected using this method in healthy NHP serum. This validation was performed entirely on frozen samples as is typical with batch assays; further testing to evaluate validation in fresh samples or samples undergoing freeze–thaw cycles are needed. Rejected cytokines were excluded from future analyses in this study since results were deemed invalid.
Matrix-M™ malaria vaccine developed by University of R21/Matrix-M™ malaria vaccine developed by University of
As we continue to work together to create a healthier, more equitable world for everyone, I am incredibly proud of the part that the Serum Institute of India has played in developing the R21 malaria vaccine. We look forward to scaling up the vaccine production to ensure that it is accessible to those who need it the most.' Weiss, S. A., Wolchok, J. D. & Sznol, M. Immunotherapy of melanoma: Facts and hopes. Clin. Cancer Res. 25(17), 5191–5201. https://doi.org/10.1158/1078-0432.Ccr-18-1550 (2019) (Epub 2019/03/30).
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To evaluate recovery (measured as observed concentration relative to expected concentration), eight unique serum pool samples were made from multiple samples taken from a single animal on different days, with the exception of one of the serum pools was constructed from samples of two different animals due to sample availability. Each sample was measured in three ways: neat (unspiked) and with spiking of a known amount of either of two manufacturer provided standards (highest concentration—kit “standard 7”—and mid-range concentration—kit “standard 5”). Each spiked sample consisted of 75% endogenous NHP serum and 25% kit standard (Supplemental Table 2). Each sample was tested in triplicate. Linearity Leng, S. X. et al. ELISA and multiplex technologies for cytokine measurement in inflammation and aging research. J. Gerontol. A Biol. Sci. Med. Sci. 63(8), 879–884. https://doi.org/10.1093/gerona/63.8.879 (2008). Liang C, Li Z, Xia X, Wang Q, Tao R, Tao Y, Xiang H, Tong S, Tao F. Determine multiple elements simultaneously in the sera of umbilical cord blood samples-a very simple method. Biol Trace Elem Res. 2017;177(1):1–8. H. Nakata, S. M. M. Nakayama, J. Yabe, K. Muzandu, H. Toyomaki, Y. B. Yohannes, A. Kataba, G. Zyambo, Y. Ikenaka, K. Choongo and M. Ishizuka, Chemosphere, 2021, 271, 129832 CrossRef CAS PubMed.
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