Blood samples for troponins I and plasma BNP levels were collected in non-pyrogenic, sterile falcon tubes. Troponin I and BNP were measured by a newly developed high-sensitive Elecsys analyzer (fully automated enzyme linked immunosorbent assay [ELISA] EVOLIS; Bio-Rad Laboratories Inc., Hercules, CA, USA). BNP kit uses competitive ELISA as the method while the cardiac-specific troponin I (cTnI) ELISA test is based on the principle of a solid phase ELISA.
Evolis
Manufactured by Bio-Rad
Sourced in United States, France, Japan
The EVOLIS is a compact and fully automated microplate processor designed for a wide range of applications in clinical and research laboratories. It features a precision liquid handling system and an integrated incubator for temperature control. The EVOLIS is capable of performing various microplate-based assays, such as ELISA and cell-based assays, with high accuracy and reproducibility.
Automatically generated - may contain errors
5 protocols using evolis
1
Quantifying Cardiac Biomarkers via ELISA
2
SARS-CoV-2 Spike RBD IgG Antibody ELISA
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Humoral immune responses were determined with a quantitative ELISA that detected anti-SARS-CoV-2-Spike-RBD IgG antibodies (Beijing Wantai Biological Pharmacy Enterprise). The ELISA was performed according to the manufacturer's instructions in a fully automated microplate processor (EVOLIS; Bio-Rad). The 32 Wantai-units per mL (U/mL) standard, part of the ELISA kit, was serially diluted to prepare six standard concentrations ranging from 32 to 1 U/mL. The standard concentrations were used to create a calibration line, whereafter antibody concentrations in serum samples were assessed. Serum IgG concentrations were converted from U/mL to international units per mL (IU/mL) using the conversion factor of 5.4 as specified by the manufacturer. Serum samples were undiluted, or if necessary, up to 1:1000 diluted to fit in the concentration range of the calibration line. HRP-conjugate was incubated at 37 °C for 30 min and TMB substrate was incubated at 37 °C for 15 min. The reaction was stopped with sulfuric acid and the absorbence was measured at 450 nm.
3
Serological Analysis of Blood Samples
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A blood sample was taken in an EDTA tube for serological analysis from each donor. The techniques used to detect the p24 antigen and anti-HIV-1 (groups M and O) and HIV-2 antibodies in donor plasma were the ELISA technique (Evolis®, BioRad, France), and the chemiluminescence technique (Cobas® 6000 e601, Roche, Germany). Serological analysis was performed according to the manufacturer's protocol.
4
Automated Immunoassay and Hematological Analysis
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The following instruments and equipment were used: Super clean bench CW-CJ-2D [Purification Equipment (Suzhou) Co., Ltd., Suzhou, China]; CO2 constant temperature incubation (Sanyo Electric Co., Ltd., Osaka, Japan); fully automatic enzyme immunoassay analyzer EVOLIS (Bio-Rad Laboratories, Inc.); fully automatic five classification hematology analyzer LH750 (Beckman Coulter, Inc., Brea, CA, USA); fully automatic biochemical analyzer DXC800 (Beckman Coulter, Inc.); electronic scale YP10002 (Shanghai Youke Equipment & Instrument Co., Ltd., Shanghai, China); electric-heated thermostatic water bath HHS-21-6 (Changzhou Nuoji Instrument Co., Ltd.); biochemical incubator DNP-9002BS-III (Shanghai Cimo Medical Devices Manufacturing Co., Ltd.).
5
Succinate Quantification in Cell Culture
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The succinate that had accumulated during cell culture was detected in the culture supernatant at the end of the incubation period according to the manufacturer's instructions (Succinate assay kit, Abcam, UK). Cell suspensions were harvested, washed and resuspended in 100 μl of ice-cold succinate assay buffer. The suspensions were centrifuged, and the collected supernatants were filtered using a 10 kDa spin filter. Fifty microliter of each sample or standard dilution were mixed with the same volume of the reaction mix or background control mix and incubated for 30 min at 37°C. The absorbance intensity was measured at 450 nm using a microplate reader (Evolis, Biorad). Each value was adjusted with the background control, in order to eliminate any non-specific absorbance that may have resulted from the possible presence of reduced nicotinamide adenine dinucleotide (NADH). Succinate concentration was then determined based on the standard curve.
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