China
Africa
Explore chapters and articles related to this topic
A Positive Correlation Between IgG2 Antibody Preponderance and Clearance of HSV During Active HSV Infections
Published in P. Mereena Luke, K. R. Dhanya, Didier Rouxel, Nandakumar Kalarikkal, Sabu Thomas, Advanced Studies in Experimental and Clinical Medicine, 2021
K. Vasanthi, G. Sathya Narayanan, Pugalendhi, Elanchezhiyan Manickan
The HSV confirmed (PCR positive) samples were classified as age-wise and gender-wise. Similar steps were done for healthy individuals to confirm HSV negativity. For the detection of total IgGs, levels (IgM, IgG) from the above serum samples EIA kits were obtained from ABCAM (Cat. No. ab137982-IgM, ab195215-IgG). For the detection of HSV, specific IgGs levels (IgM, IgG) from the above serum samples EIA kits were procured from CALBIOTECH, USA, and MYBIOSOURCE (USA) (Cat. No. H1M4908, H2M4989, H1G5010, H2G4881). For the detection of IgG subclasses from the above demography, the EIA kits were obtained from Invitrogen-ThermoScientific (Cat. No. 991000). The analysis of immunoglobulins levels on serum samples was performed according to the manufactures instructions. Then the plates were read at 450 nm in an ELISA plate reader. A standard curve was plotted by linear regression analysis using the values of the standard and the values of the unknown samples were calculated from the graph.
Analytical Method and Assay
Published in Emmanuel Lesaffre, Gianluca Baio, Bruno Boulanger, Bayesian Methods in Pharmaceutical Research, 2020
Ligand-binding assays, in particular ELISA, are widely used to quantify proteins. The mechanism of detection is based on biology by opposition as for chromatographic methods relying on physico-chemical properties. Quantification of the amount of analyte existing in samples is usually performed by relating the signal of the assay (optical density) to concentration of a standard sample, namely a calibration curve. Typical models used to fit the standard curve are non-linear, such as 4 or 5-parameter logistic functions (see Figure 20.6). Numerous assay parameters can be tuned to optimize such assays such as the nature of enzymes, volumes of products, different incubation times, etc. (Yarovoi et al., 2013; Verch, 2014). Each combination of these parameters will influence the shape of the calibration curve and hence alter the predicted concentration of analyte. The final aim of such assays is to be able to provide accurate analytical results. This is generally translated in an operational way by requiring having results with less than λ% total error, e.g. 30%. To this end, the path to optimization is to select the ranges of the assay parameters that will reach high probability to provide accurate results.
Diagnostic applications of immunology
Published in Gabriel Virella, Medical Immunology, 2019
Ajay Grover, Virginia Litwin, Gabriel Virella
The enzyme-linked immunosorbent assay (ELISA), currently known as enzyme immunoassay (EIA), is a flexible platform for the detection of an unlimited number of antigens in a variety of matrices. EIA is most often performed in a 96-well plate format and can be highly specific and sensitive in the nanomolar and picomolar ranges, when the correct configuration of high-quality, highly specific, serological reagents (monoclonal or polyclonal) is incorporated into the assay. The assay can be quantitative if an appropriate standard curve is incorporated into it. With the use of the 96-well plate format and automated pipetting devices, EIA can be high throughput and can be performed in different configurations:
M6P-modified solid lipid nanoparticles loaded with matrine for the treatment of fibrotic liver
Published in Drug Delivery, 2023
Xiaochuan Tan, Yumei Hao, Nai Ma, Yige Yang, Wenzhen Jin, Ya Meng, Chuchu Zhou, Wensheng Zheng, Yujia Zhang
A total of 180 C57BL mice were randomly divided into five groups (each group of 32 mice) and administered the corresponding drugs: control group (healthy mice without CCl4), Model group (CCl4 but no drug), MT-solution group (CCl4 with free MT solution), MT-SLN group (CCl4 with MT-SLN), and M6P-HSA-MT-SLN group (CCl4 with M6P-HSA-MT-SLN). Mice were injected intravenously with corresponding MT solution (20 mg/kg) via the tail three times a week. The serum and liver samples of 8 mice in each group were collected once a week. And blood samples were collected from each mouse at the end of 4 weeks, then the level of AST, ALT, HA, LN, PC III, and Col IV in serum were measured. The body weight was also measured and recorded. The level of liver fibrosis biomarkers (AST, ALT, HA, LN, PC III, and Col IV) in the serum of each mouse was compared before and after drug administration using the ELISA kits. The standard curve was produced according to the operation instructions, and the concentration of biomarkers was determined according to the standard curve. Hematoxylin-eosin (H&E), Masson staining, and Sirius Red staining were utilized to measure the degree of liver fibrosis in mice to calculate the effect of the M6P-HSA-MT-SLN. The image analysis software Pro-Plus 7.0 was used to estimate the proportion of inflammation/necrotic lesion involvement area in the total liver tissue section. And the IHC of α-SMA and collagen I of liver sections was used to investigate antifibrotic effect on hepatic stellate cells of different MT formulations.
Pharmacokinetic and pharmacodynamic modeling of gut hormone peptide YY(3–36) after pulmonary delivery
Published in Drug Development and Industrial Pharmacy, 2019
Jie Shao, Mong-Jen Chen, Philip J. Kuehl, Guenther Hochhaus
In brief, aqueous solutions of human PYY(3–36) were administered to C57BL/6 mice through intraperitoneal (IP) injection (0.1 mg/kg) or subcutaneous (SC) injections (0.03, 0.1, 0.3 mg/kg) to assess the pharmacokinetics after parenteral administration. For pulmonary delivery, mice inhaled different doses of PYY(3–36) dry powder formulations via a nose-only inhalation exposure system, resulting in pulmonary deposited doses of 0.006, 0.02, 0.21 mg/kg, determined as previously described [19]. Over the investigation period of 6 h, mice (n = 3 at each time point) were euthanized at defined time points and blood was collected via cardiac puncture. PYY(3–36) concentrations were determined using a commercial available enzyme-linked immunosorbent assay (ELISA) kit (Millipore Biopharma Services, Billerica, MA, limit of quantification of 0.05 ng/mL). The precision and accuracy of the standard curve were reported to be within ±25% of nominal concentrations in the range of 0.05–50 ng/mL.
Cryo-thermal therapy inducing MI macrophage polarization created CXCL10 and IL-6-rich pro-inflammatory environment for CD4+ T cell-mediated anti-tumor immunity
Published in International Journal of Hyperthermia, 2019
Ping Liu, Shengguo Jia, Yue Lou, Kun He, Lisa X. Xu
To evaluate the level of CXCL10 and IL-6 in spleen after cryo-thermal therapy, the spleen tissues were collected from cryo-thermal-treated mice or hyperthermia-treated mice on day 14 after the treatment, and tumor-bearing mice on day 26 after tumor inoculation, and tissue homogenate was prepared using 1 × PBS. To evaluate the level of CXCL10 and IL-6 in serum after cryo-thermal therapy mice, the peripheral blood was collected from cryo-thermal-treated mice or hyperthermia-treated on day 14 after the treatment and tumor-bearing mice on day 26 after tumor inoculation. The peripheral blood samples were centrifuged and the serum was obtained. Then the CXCL10 and IL-6 concentration in the supernatant of spleen tissue homogenate and serum were analyzed using mouse CXCL10/IP-10 ELISA kit and mouse IL-6 ELISA kit (all from Boster, Pleasanton, CA), respectively. A standard curve was established according to the manufacture’s instruction. Experimental values were computed with the use of regression analysis.