His-Express Detection EIA Kit

Wastewater production has also increased due to increasing population growth and increasing water consumption in communities. This increase in sewage production can be a major environmental hazard. Hospital wastewater is one of the types of wastewater that poses many hazards to the environment and is classified as hazardous by its quality. Therefore, essential to the output status hospital wastewater should be accurate and prevent hospital wastewater from entering without purifying the living environment. The aim of this study was the design and synthesis construct containing the lipase-subtilisin fusion gene in the plasmid pet22b vector. The recombinant plasmid then was transformed and expressed in expression host E. coli BL21DE3. Construct PET22b-LIP-SUB was designed by researchers and optimized with bioinformatics software. It was then synthesized by a Biometrics company. The transformation was performed and was induced in the presence of 1 mM concentration IPTG. For confirmation of recombinant protein SDS-PAGE and western blotting were performed. Nickel column was used for purification of the recombinant protein. Based on bioinformatics analysis, the recombinant protein had good spatial structure and stability and half-life. The recombinant protein was confirmed by SDS-PAGE and western blotting with Anti-his tag. The concentration of the recombinant protein was 4.11 mg/ml. The results of this research showed that recombination protein LIP-SUB contains heat and acid resistant and higher potency, therefore, require to evaluate biological activity in hospital wastewater.

A helical epitope-peptide (lle(85)-Gly(94)) was selected from the alpha-helix structure of the HIV protease (PR) as the template, which represents an intricate interplay between structure conformation and dimerization. The peptide template was mixed with water, trifluoroethanol (TFE), and acetonitrile (ACN) at a certain ratio to enlarge the helical conformation in the solution for the fabrication of helical epitope-mediated molecularly imprinted polymers (HEMIPs) on a quartz crystal microbalance (QCM) chip. The template molecules were then removed under equilibrium batch rebinding conditions involving 5% acetic acid/water. The resulting HEMIPs chip exhibited a high affinity toward template peptide HIV PR85-94, His-tagged HIV PR, and HIV PR, with dissociation constants (K-d) as 160, 43.3, and 78.5 pM, respectively. The detection limit of the developed HIV PR(85-94)QCM sensor is 0.1 ng/mL. The HEMIPs chip exhibited a high affinity and selectivity to bind HIV PR and subsequently to an inhibitor of HIV PR (nelfinavir). The HIV PR binding site was properly oriented on the HEMIPs-chip to develop a HIV PR/HEMIPs chip, which can effectively bind nelfinavir to establish a sandwich assay. The nelfinavir then attached to the HIV PR/HEMIPs chip, which can be easily removed involving 0.8% acetic acid/water. Therefore, HIV PR/HEMIPs chip can be useful to screen for other HIV PR inhibitors. This technique may improve drug targeting for HIV therapy and also strengthen investigations into other virus assays.