Screening venous blood samples were analyzed for parasitemia and biochemistry (point-of-care iSTAT analyzer [CG4+, Chem8+], Abbott Laboratories, USA). After enrollment, vital signs, urine output, development of complications, and parasitemia were assessed every 6 hours until discharge or death. For the first 72 hours, serum creatinine was assessed every 12 hours (Olympus AU400 chemistry analyzer, performed in Bangkok), CFH in twice-centrifuged citrated plasma was assessed every 24 hours (enzyme-linked immunosorbent assay; Bethyl Laboratories, performed in Darwin) [9 (link)], and F2-IsoPs and IsoFs in lithium heparin plasma were assessed every 24 hours (gas chromatography-mass spectrometry at Vanderbilt University) [7 (link), 8 (link)]. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were assessed on enrollment and at 72 hours. To determine the PK–PD of acetaminophen on creatinine, parasitemia, and fever, plasma EDTA samples for acetaminophen concentration were collected prior to each dose plus dense sampling in the treatment arm after both the first (0 hour) and last (72 hour) dose. Patients in the control arm had samples collected every 6 hour for 72 hours to assess unplanned acetaminophen intake. Detailed procedures are provided in the Supplementary Material .
Enzyme linked immunosorbent assay
Manufactured by Fortis Life Sciences
Sourced in United States
The Enzyme-linked immunosorbent assay (ELISA) is a laboratory technique used to detect and quantify specific proteins, antibodies, or other molecules in a sample. It is a sensitive and specific analytical method that utilizes antigen-antibody interactions and enzyme-catalyzed color changes to measure the presence and concentration of target analytes.
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Lab products found in correlation
Blood glucose meter, by Roche (2 mentions)
Au400 chemistry analyzer, by Olympus (1 mentions)
Fuji dri chem, by Fujifilm (1 mentions)
Architect c8000 chemistry analyzer, by Abbott (1 mentions)
Clinitek 50, by Bayer (1 mentions)
Urea nitrogen test, by EKF Diagnostics (1 mentions)
Spectramax i3 plate reader, by Molecular Devices (1 mentions)
Metabolic cage, by Tecniplast (1 mentions)
Premier hb9210 hba1c, by Trinity Biotech (1 mentions)
Automatic biochemical analyzer, by Roche (1 mentions)
13 protocols using enzyme linked immunosorbent assay
1
Acetaminophen PK-PD in Malaria Patients
2
Quantifying Human Albumin in Rat Blood
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Whole blood samples were centrifuged (4000 revolutions per min) for 20 min at 4 °C, and the supernatants were collected and stored at −30 °C. Subsequently, the supernatants were tested using a serum multiple biochemical analyzer (Fuji Drichem; Fuji Film Inc., Tokyo, Japan). The human albumin level in the rat blood was assessed by enzyme linked immunosorbent assay (Bethyl Laboratories, Montgomery, TX, United States), according to the manufacturer’s instructions.
3
Comprehensive Monitoring of Malaria Complications
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Venous blood was collected on enrollment for standard hematology and biochemistry. Peripheral blood parasitemia was assessed by research microscopists on enrollment and every 6 hours until 2 consecutive negative smears were obtained. Serum creatinine was measured on enrollment and then every 12 hours until 72 hours, then at 7 and 28 days. Creatinine was measured in real time at the accredited enrolling hospitals using an automated modified Jaffe alkaline picrate method (Architect c8000 Chemistry Analyzer, Abbot), with the assay traceable to the Isotope dilution mass spectrometry (IDMS) standard. Urine microalbumin was measured on fresh midstream urine (Clinitek 50, Bayer). CFHb was measured at enrollment, at 12 hours, and then daily for 72 hours using enzyme-linked immunosorbent assay (Bethyl Laboratories) on twice-centrifuged citrated plasma. Liver function tests (LFTs) and urine albumin-to-creatinine ratio (ACR) were measured at 0 and 72 hours and at 7 and 28 days. Plasma acetaminophen concentrations were measured using liquid chromatography-mass spectrometry as previously described [26 (link)], detailed in the Supplementary Materials .
4
Characterization of Encapsulated hPSC-Heps
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The encapsulated hPSC-Heps were characterized by albumin secretion and ureagenesis to ensure that the encapsulation process did not abolish hepatocyte functions. Albumin secretion was measured by an Enzyme-linked immunosorbent assay using a commercially available kit (Bethyl Laboratories) (n = 5). Media from encapsulated cells were collected at 5 and 14 days post-encapsulation. Ureagenesis was measured by a commercially available colorimetric assay kit (QuantiChrom). The encapsulated cells were challenged by 4 mM NH4Cl in the culture medium for 24 h, and media supernatant samples were collected and analyzed for urea content subsequently (n = 3).
5
Quantification of Human Albumin in Mice
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Quantification of human albumin was carried out as described previously [21 ]. All blood samples of the mice were collected before transplantation and then were collected every week after transplantation. Blood samples were centrifuged by 4000 rpm, 20 min at 4 °C, and the plasma was collected. The collected plasma was diluted, and the human albumin concentrations were assessed by enzyme-linked immunosorbent assay (Bethyl Laboratories, Montgomery, TX, USA), according to the manufacturer’s instructions.
6
Liver MPS Functional Characterization
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Perfusion of the liver MPS was initiated 2 days after loading to allow recovery and spreading of the hiPSC-Heps. Media was collected every two days and analyzed for albumin and urea secretion. Albumin was measured using an enzyme-linked immunosorbent assay (Bethyl Laboratories). Urea nitrogen was measured using a colorimetric assay (urea nitrogen test, Stanbio Laboratory) modified to be performed in a 384-well microtiter plate, including increase in the incubation time of reactants from 60 to 90 min at 60°C prior to reading. All the biochemical assays were performed in 10 µL of media and measured on a SpectraMax i3 plate reader (Molecular Devices). All the sample results were calculated by interpolation of sample raw values from standard curves performed in parallel. The same assays were repeated for conventional cell cultures of hiPSC-Heps.
7
Quantification of Human Albumin in Mice
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Quantification of human albumin (ALB) was carried out as described previously [25 (link)]. All blood samples of the mice were collected before transplantation and then collected every week after transplantation. Blood samples were centrifuged by 4000 rpm, 20 min at 4 °C, and the plasma was collected. The collected plasma was diluted, and the human albumin concentrations were assessed by enzyme-linked immunosorbent assay (Bethyl Laboratories, Montgomery, TX, USA), according to the manufacturer’s instructions.
8
Fasting Glucose and Urinary Albumin Assay
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The level of fasting blood glucose of all mice was monitored 4 h post feeding by using blood glucose meter (Roche, Basel, Switzerland). Urinary albumin excretion was detected by the enzyme-linked immunosorbent assay (Bethyl Laboratories, Montgomery, United States) according to the manufacturer’s instructions.
9
Limiting Dilution Assay for Tumor-Initiating Cells
10
Urine Albumin Measurement in Celsr1 Mice
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Urine was collected from 3-month-old Celsr1Crsh/+ and wild-type littermates by housing them individually in metabolic cages. Albumin concentrations were measured by enzyme-linked immunosorbent assay (Bethyl Laboratories, Montgomery, Alabama).53 (link)
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