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HLA-DR and -DQ Serotyping
Published in M. Kam, Jeffrey L. Bidwell, Handbook of HLA TYPING TECHNIQUES, 2020
The majority of the useful monoclonal antibodies presently available have been generated using antibody-producing splenocytes from mice immunized with human cells. Some antibodies with good anti-HLA activity, have been produced as a result of immunizing rats,11 and techniques have also been developed to transform and immortalize B lymphocytes from antibody-producing human subjects to make monoclonal antibodies.12Table 4 lists a number of cytotoxic monoclonal antibodies with anti-HLA-DR and -DQ reactivity. Comparatively few useful monoclonal antibodies are available for the definition of the DRB1 HLA-DR specificities. This may be related to the nature of the response of the mouse immune system to human DR molecules. However, a number of reagents help to define certain specificities, such as DR103, DR12, and DR14, for which few sera are available. Monoclonal antibodies with anti-DR52 activity are also useful as it is known that they do not have any activity toward DRB1-encoded molecules.13 Monoclonal antibodies are most useful for the definition of HLA-DQ specificities as they are known to be devoid of anti-HLA-DR activity. The anti-DQ4 monoclonal antibody listed in Table 4 was the first reagent to clearly define this specificity,14 and the antibody ′TIBs’′ can be used to identify the presence of DQ8 or DQ9 in the presence of DQ7.15 This is not possible using antisera.
Emerging Potential of In Vitro Diagnostic Devices: Applications and Current Status
Published in Debarshi Kar Mahapatra, Sanjay Kumar Bharti, Medicinal Chemistry with Pharmaceutical Product Development, 2019
Swarnali Das Paul, Gunjan Jeswani
According to the definition of In vitro diagnostic/medical device they are basically “a device, whether used alone or in combination, is specimen derived from the human body solely or principally to provide information for diagnostic, monitoring or compatibility purpose.” They comprise of calibrators, reagents, software, specimen, receptacles, control materials, instruments or other items according to USFDA [1]. Here reagent signifies as biological, chemical, or immunological articles, proposed by the manufacturer. In vitro diagnostic (IVD) devices also include the genetic investigations which reveal information for healthcare decision making [2, 3]. An IVD may be either an entire test or a part of a test. Part of the test includes both non-diagnostic components, known as “general purpose reagents” (GPRs), and also the active element of the diagnostic test, called as “analyte specific reagent” (ASR). IVDs which are used in the clinical monitoring of patients also referred as a medical device. Thus, due to their critical requirements, they are subjected to regulation by the FDA.
Principles of Clinical Pathology
Published in Pritam S. Sahota, James A. Popp, Jerry F. Hardisty, Chirukandath Gopinath, Page R. Bouchard, Toxicologic Pathology, 2018
Niraj K. Tripathi, Jacqueline M. Tarrant
Reagent strip tests are only semiquantitative measures, and care must be taken to avoid overinterpretation of results. Animals with highly concentrated urine often appear to have a higher incidence of positive results, or greater amounts of an analyte, than animals with dilute urine simply because of urine concentration. A more quantitative measurement normalized for concentration may be desirable for proper interpretation of some test results (e.g., increased urine protein). Results from automated reagent strip readers using a gray-scale system can also be affected by urine color, and manual examination of reagent strips may be warranted if the test article or a metabolite causes an abnormal color.
Group 2 innate lymphoid cells boost CD8+ T-cell activation in anti-tumor immune responses
Published in OncoImmunology, 2023
Jing Wen, Shipeng Cheng, Ran Wang, Yuying Huang, Long Xu, Liyan Ma, Zhiyang Ling, Jinfu Xu, Deping Zhao, Yaguang Zhang, Bing Sun
The LDH Cytotoxicity Assay Kit was purchased from Beyotime (C0016, Beyotime). Experiments were performed according to the manufacturer’s protocol. Briefly, 1 × 104 B16F10-OVA tumor cells were seeded in 48-well plates 1 day prior to establishing the co-culture system. A total of 1 × 105 OT-I CD8+ T cells mixed with ILC2s or ILC2s previously loaded with OVA were added to adherent tumor cells. Simultaneously, OT-I CD8+ T cells alone, ILC2s alone, or previously OVA-loaded ILC2s alone were added to the tumor cell culture system as controls. After 48 h, the culture supernatant was aspirated from each well. The reagents in the kit were successively added to the test samples as described in the protocols. The absorbance of all test samples was measured using a 450-nm filter. The results were used to calculate LDH release from tumor cells using a standard curve. The relative value was calculated based the formula: (Absorbance of sample – Absorbance of control sample)/(Absorbance of the maximum enzyme activity of the cell-Absorbance of control sample) × 100%。
Every nano-step counts: a critical reflection on do’s and don’ts in researching nanomedicines for retinal gene therapy
Published in Expert Opinion on Drug Delivery, 2023
Karen Peynshaert, Joke Devoldere, Stefaan De Smedt, Katrien Remaut
Along with the entire biomedical field, the landscape of nano delivery is contorted by a severe reproducibility crisis[77]. Shockingly, it is estimated that the total prevalence of irreproducible preclinical studies exceeds 50%. It goes without saying that this lack of reproducibility is extremely inefficient and dramatically slows down any advancement to the clinic[78]. Sadly, this also implies that a tremendous amount of funding is spent on research that cannot be replicated which is for the US alone estimated around 28 billion dollars per year. An extensive analysis by Freedman et al. indicates that the primary roots of irreproducibility are 1) study design, 2) biological reagents and reference materials, 3) laboratory protocols, and 4) data analysis and reporting[79]. On top of these key points, Voelkl et al. further argues that the consistent ignorance of the biological variation among animals within the study design is a dominant cause[77].
Flavonoid-rich fraction of Lasianthera africana leaves alleviates hepatotoxicity induced by carbon tetrachloride in Wistar rats
Published in Drug and Chemical Toxicology, 2022
Daniel Emmanuel Ekpo, Parker Elijah Joshua, Arome Solomon Odiba, Okwesilieze Fred Chiletugo Nwodo
All chemicals and reagents used for the study were of analytical grade, and products of British Drug House (BDH), England (chloroform; methanol; ethanol; ammonia; trichloroacetic acid, TCA; trioxonitrate (V), sulfuric acid, and hydrochloric acid), May and Baker, England [sodium chloride and sodium trioxcarbonate (IV)], Qualikems, India (aluminum chloride, potassium acetate, and phosphomolybdic acid) and Sigma Aldrich, St. Louis, MO (quercetin, gallic acid, sodium hydroxide, ferric chloride, ethyl acetate, acetic anhydride, DOWEX ion-exchange resin). Reagent kits used for biochemical assays were purchased from Randox Laboratories Ltd., Crumlin, UK. Glass distilled and deionized water were obtained from the National Center for Energy Research and Development (NCERD), University of Nigeria, Nsukka.