ABSTRACT
Background: Dexamethasone is a synthetic corticosteroid similar to cortisol produced naturally by the adrenal glands. As an anti- inflammatory and immunosuppressive agent, it is used in many diseases such as rheumatoid arthritis and allergic anaphylactic shock, and its suppression test to diagnose Cushing's syndrome. Its further use includes its administration before antibiotics in bacterial meningitis, antitumor treatment, for treatment of glucocorticoid resistance, Addison抯 disease, and congenital adrenal hyperplasia. The drug is abused by using it in animal husbandry as a growth promoter and in horse sports to enhance their performance. Methods: In this study, the development of homologous ELISA using Dexamethasone-21-hemisuccinate (DEX-21-HS)-Bovine serum albumin antiserum and Dexamethasone-21-hemisuccinate (DEX-21-HS)-Horseradish peroxidase enzyme conjugate has been done. The n-hydroxysuccinimide ester method was used to prepare the immunogen and enzyme conjugate. Results: The sensitivity 0.25 ng/mL, affinity 2.8x10-8 L/mol and ED50 4.98 ng/mL of the assay were found. The cross-reactivity of the assay was checked and found with three steroids (Corticosterone- 1.13%, Progesterone- 2.25% and Prednisolone- 6.3%) out of 48 structurally related steroids. Then, analytical variables of the developed assay were studied, such as recovery (98.55% to 105.08%), precision (Inter and Intra- assay coefficient of variation <9.28%), correlation (R2= 0.98) by utilizing a commercially available Dexamethasone kit for comparison. Conclusion: This study concluded that low-cost indigenous ELISA for Dexamethasone had been developed, which can give results within 75-80 minutes.
ABSTRACT
Objective: To obtain the monoclonal antibodies against the protein of ginkgo diterpene lactones meglumine injection. Methods: After total proteins of ginkgo leaf running two-dimensional electrophoresis, the protein spots were digged and analyzed by mass spectrometry (MS). According to the result of MS, the polypeptide was synthesized and used to immunize the BALB/c mice. The indirect ELISA was utilized to develop monoclonal antibody after cell fusion between SP2/0 cells and spleen cells from the immunized BALB/c mice. Results: The immunoglobulin subtypes of five monoclonal antibodies were IgM and light chain was Kappa identified with a commercial capture-ELISA kit. Western blotting analysis also indicated that the monoclonal antibodies were specific to the protein of ginkgo leaf. Conclusion: The experiment would be helpful for the establishment of detection method specific to the proteins of ginkgo diterpene lactones meglumine injection in the future study.
ABSTRACT
We have generated a chimera between the enzymatically active A subunit of the E coli derived AB5 verotoxin and a single receptor-binding B subunit. The construct was made by in frame fusion of the 3’ terminus of the A subunit gene with the 5’ end of the B subunit gene via the deletion of the intervening bases of the verotoxin operon such that the C terminus of the A subunit is continuous with the N terminus of a single B subunit. The gene product is a single fusion protein of 38kDa molecular weight, reactive with polyclonal and monoclonal antibodies against either the A or B subunits of the wild type toxin. The chimera showed a 104-105 fold reduction in cell vero cell cytotoxicity but no toxicity for the globotriaosyl ceramide (Gb3) deficient VRP subclone. No Gb3 binding by TLC overlay was detected. Polyclonal rabbit anti-VT1A-B chimera serum neutralizes VT1 cytotoxicity in vitro but reacts only with the A subunit of the wildtype holotoxin by western blot. This A-B chimera illustrates the importance of the pentameric B subunit in receptor binding and potentially identifies a novel attenuation vaccination strategy.
ABSTRACT
The need for an efficacious HIV/AIDS vaccine remains the highest priority of the world HIV/AIDS agenda. The generation of an efficacious HIV/AIDS vaccine proves an enormous scientific challenge. This article reviews the neutralizing antibody problem, elusive immune protection, immunogen design, pre-existing anti-vector immunity and design of phase 3 vaccine trials and the challenges and opportunities in development of HIV/AIDS vaccine are discussed.
ABSTRACT
Objective:To prepare anti-H pylori immune milk to prevent the infection of H pylori.Methods:The whole active H pylori as immunogen, cows were immunized and the polyclonal immune milk to H pylori was obtained. Results: With direct agglutination test and doubled agar diffusion detection, the titer of antibody was 1:2 048 and 1:32.Conclusion: The prepared immune milk against H pylori has good polyclonal specificity and high antibody titeration.