To determine the safety of the VLP vaccine candidates and their implant formulations, especially focusing on the trivalent formulation, toxicity plasma biomarkers were assessed. For liver damage the concentrations of the enzymes aspartate transaminase (AST) and alanine transaminase (ALT) were established by Aspartate Aminotransferase Activity kit [Abcam] and Alanine Transaminase Activity Assay Kit [Abcam], respectively. For kidney damage the concentration of kidney injury molecule-1 (KIM-1) by Mouse KIM-1 ELISA Kit [Abcam] was determined. Here, plasma samples collected at week 0 and 12 weeks post-immunization were tested. The plasma samples were evaluated per animal or pooled by group and tested in triplicate.
Mouse kim 1 elisa kit
Manufactured by Abcam
Sourced in United Kingdom
The Mouse KIM-1 ELISA Kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of mouse kidney injury molecule-1 (KIM-1) in biological samples.
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Lab products found in correlation
Alanine transaminase activity assay kit, by Abcam (1 mentions)
Easy protein quantitative kit, by Transgene (1 mentions)
Balb c mice, by Charles River Laboratories (1 mentions)
Triglyceride reagent gpo pap, by Roche (1 mentions)
Ultra sensitive mouse insulin elisa, by Crystal Chem (1 mentions)
Activity assay kits, by Abcam (1 mentions)
Protease inhibitor cocktail, by Nacalai Tesque (1 mentions)
Infinite m200 pro, by Tecan (1 mentions)
6 protocols using mouse kim 1 elisa kit
1
Toxicity Biomarkers for VLP Vaccine Candidates
2
Pristane-Induced Lupus Model in Mice
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Six- eight weeks old wild-type female BALB/c mice were obtained from Beijing Vital River Laboratory Animal Technology Co., Ltd. Before induced by pristane i.p (500 μl/mouse), mice (10 mice in each group) were immunized with KLH-HU1 or KLH every 3 weeks for a total of six times. For each mouse, urine and serum samples were collected every two weeks. Urine total protein was determined by Easy Protein Quantitative Kit (TransGen Biotech). The levels of urinary KIM-1 (kidney injury molecule-1) were measured by Mouse KIM-1 ELISA Kit (ab213477, Abcam). The anti-dsDNA antibodies in the serum were quantified with anti-dsDNA IgG ELISA kit (EUROIMMUN). All mice were sacrificed at week 30 after pristane injection, and the kidney were obtained to take pathological examination by HE staining and immune complex detection by fluorescence microscopy.
3
Biomarkers of Metabolic Disorders
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The plasma levels of triglyceride (TG) were measured using Roche triglyceride reagent GPO-PAP (Roche Life Science) and the levels of non-esterified fatty acid (NEFA) were measured using Wako HR Series NEFA-HR(2) kit (Fujifilm, Tokyo, Japan). Blood insulin and urinary albumin concentration were measured using Ultra-Sensitive Mouse Insulin ELISA and Mouse Albumin ELISA Kits (CrystalChem, Elk Grove Village, IL, USA), respectively. Urinary KIM-1 was measured using a Mouse KIM-1 ELISA Kit (Abcam, Cambridge, UK). The concentrations of urinary and blood creatinine were determined using Urinary and Blood Creatinine Assays, respectively (Cayman Chemicals, Ann Arbor, MI, USA). All assays were performed according to the manufacturers’ instructions.
4
Assessing QβS100A9 Vaccine Safety
5
Kidney Injury Biomarker Quantification
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Kidney tissues were homogenized with radioimmunoprecipitation (RIPA) buffer lysis buffer (Wako, Osaka, Japan) containing 1% protease inhibitor cocktail (Nacalai Tesque), and the supernatant was used for measuring kidney injury molecule-1 (KIM-1; Mouse KIM-1 ELISA Kit, Abcam) and neutrophil gelatinase-associated lipocalin (NGAL; NGAL ELISA Kit, Enzo Life Science, New York, NY). Urinary KIM-1 levels were also measured and normalized by urine creatinine level (Dry Chem 3500 V, Fujifilum, Tokyo, Japan).
6
Quantifying Urinary KIM-1 for Nephrotoxicity
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Among a panel of urinary biomarkers, KIM-1 has been shown to be best suited for detecting OTA-induced nephrotoxicity (Hoffmann et al. 2010 (link)). Concentrations of KIM-1 were determined in urine using the Mouse KIM 1 ELISA Kit from Abcam (ab213477) according to manufacturer’s protocol. Briefly, the previously diluted urine samples and mouse recombinant KIM-1 standards were added to the wells followed by the antibody cocktail (capture and detector antibodies) and incubated for 1 h at room temperature on a plate shaker. After the incubation, the antibody-KIM-1 sandwich complex was monitored by streptavidin conjugated to horseradish peroxidase (HRP). Finally, the optical density of the color-forming TMB substrate was measured at an optical density of 450 nm using a microplate reader (Infinite M200 Pro, Tecan) and the concentration of KIM-1 of each sample was calculated from the standard curve.
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