Quantitation of Biomarkers in Clinical Samples

IL1-α, IL-8, and VEGF-A in cell supernatant were analyzed using Quantikine ELISA kits (R&D Systems; DLA50, D8000C, DVE00). Human GRO-beta and GRO-gamma ELISA Construction kits (Antigenix America Inc.; RHF810CK, RHF820CKP) were used to measure the supernatant and plasma levels of CXCL2 and CXCL3, respectively. IL-1α, IL-8, and VEGF-A in human plasma were analyzed using custom V-PLEX Assays (Meso Scale Discovery, Maryland, USA; K151RBD-2, K151RAG-1, K151RHD-1). Patient blood samples were obtained after multicenter ethics approval (09/H1102/107 and 14/YH/0090) as described previously [16 (link), 22 (link)]. Blood was centrifuged within 1 h of collection at 2500 rpm in a lithium-heparin containing tube; the plasma was then removed, aliquoted, and stored in a − 70 °C freezer prior to thawing for ELISA analysis. ELISAs were carried out according to the manufacturer’s instructions.

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Phillips M.M., Pavlyk I., Allen M., Ghazaly E., Cutts R., Carpentier J., Berry J.S., Nattress C., Feng S., Hallden G., Chelala C., Bomalaski J., Steele J., Sheaff M., Balkwill F, & Szlosarek P.W. (2023). A role for macrophages under cytokine control in mediating resistance to ADI-PEG20 (pegargiminase) in ASS1-deficient mesothelioma. Pharmacological Reports, 75(3), 570-584.

Publication 2023

Quantikine ELISA kits DLA50 V-PLEX Assays

Assays Blood Cell Elisas Gro beta Gro gamma Heparin Human Il1 α Lithium Patient Plasma Vegf

Corresponding Organization :

Other organizations : Queen Mary University of London, Cancer Research UK, Institute of Cancer Research, Breast Cancer Now, Barts Health NHS Trust, St Bartholomew's Hospital, Royal London Hospital

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

IL1-α, IL-8, and VEGF-A in cell supernatant
CXCL2 and CXCL3 in supernatant and plasma
IL-1α, IL-8, and VEGF-A in human plasma

control variables

Patient blood samples were obtained after multicenter ethics approval (09/H1102/107 and 14/YH/0090) as described previously [16, 22]
Blood was centrifuged within 1 h of collection at 2500 rpm in a lithium-heparin containing tube; the plasma was then removed, aliquoted, and stored in a − 70 °C freezer prior to thawing for ELISA analysis

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