Preparation, Purification and Performance Evaluation of Polyclonal Antibody Against SARS-CoV-2 Produced in Rat

Purpose: Among all known human coronaviruses, some viruses (e.g., SARS-CoV-2) cause severe pneumonia or even death. With the regard to its spread and the importance of its rapid identification/treatment, and because pAbs are relatively cheap, able to bind to more sites on antigens and even neutralize them, this study was done for the production and purification of anti-SARS-CoV-2 polyclonal antibodies (pAb) in rats. Methods: Viral antigen purification was performed by PEG/NaCl precipitation. The efficiency of the sucrose cushion method was also investigated to produce a purer antigen. Immunization was done and antibody purification was performed by ammonium sulfate precipitation (33%), dialysis, and ion-exchange chromatography. Western blotting and enzyme-linked immunosorbent assay (ELISA) were performed to verify the antibody specificity. All data were analyzed by SPSS software. Results: The results showed that the amount of concentrated virus increased with the increase of PEG concentration. Moreover, the sucrose cushion method increased its purity. Besides, induction of immune response in rats for pAb production with high titers was reached via these antigens and ELISA/western blot results indicated a suitable antibody-antigen interaction. Additionally, it was shown that ion-exchange chromatography could be a suitable technique for IgG purification. Conclusion: Herein, we presented a simple and cheap method for the purification of whole viral particles with relatively high quality. The results verified that these antigens could elicit a good immune response in the rat. The obtained pAbs showed a good specificity toward SARS-CoV-2 antigens. Accordingly, this study proposes a promising method for viral vaccine development

Screening and identification of efficient strain in selenium oxyanions sorption in order to biological wastewater treatment

Introduction: Selenium is an element with antioxidant activities that plays roles in thyroid hormone homeostasis, immunity and also fertility. Nevertheless, selenium toxicity (selenosis) causes problems for humans such as abnormalities of the nervous system, gastrointestinal problems and hair loss. Thus, this study was performed with the aim of bacterial biosorbent isolation in order to remove selenium contaminant from wastewater. Materials and methods: In this research, at first using modified Luria- Bertani agar (mLBA) medium with certain concentration of sodium selenate salt, isolation of bacterial isolates was done from three collected wastewater and sludge samples from Khouzestan industrial factories. After determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), the sorption capacity and the percentage of metal removal efficiency (%RE) were investigated by atomic absorption spectrophotometer using metabolically active and inactive samples belonging to an efficient isolate. Identification was performed by morphological, biochemical and molecular methods. Results: Among 73 attained bacterial isolates at the first stage, 8 selenate oxyanion resistant isolates were gathered. Among these, AMS1-S8 isolate with MIC= 600­mM and MBC= 1200­mM were selected for more studies. Attained results in sorption mechanism determination stage showed that the sorption capacity in metabolically active sample is more than the inactive samples. Based on the identification results, it is revealed that this isolate belongs to the Enterobacter genus. This isolate is deposited as accession JQ965667 in the GeneBank database. Discussion and conclusion: The results showed that active biomass of selected isolate, have most sorption capacity and %RE and among the other isolates, have high partial resistance against selenate. Therefore, it can be a relatively ideal option for the bioremediation of polluted environments