ABSTRACT
@#Objective: To evaluate the sensitivity and the stability of the monoclonal antibodies (Aa3c10, b10c1), against truncated Histidine-rich protein 2 (PfHRP2), developed using smart polymer, poly N-isopropylacrylamide, as adjuvant for malarial diagnostic applications in comparison with the available commercial antibodies. Methods: Two hybridoma clones (Aa3c10, b10c1) were used for the production of ascites in BALB/c mice. Purification of monoclonal antibodies from the ascites was carried out using affinity columns. The thermal stability study of monoclonal antibodies was done by storing it at 37°C and 45°C for thirty days. The stored antibodies were analyzed using SDS-PAGE and flow-through device where the antigenantibody interaction was visualized by Protein A colloidal gold solution. Sensitivity was determined by endpoint dilution ELISA and the dissociation constant by competitive ELISA. Sensitive pair optimization was done by sandwich ELISA using biotinylated antibodies. Prototype preparation for lateral flow assay had a colloidal gold-based detection system. Results: Thermal stability experiments showed that both mAbs (Aa3c10; b10c1) are stable up to thirty days at 45°C while the commercially available mAbs were stable up to fifteen days only. Compared to commercial antibodies, the mAb Aa3c10, showed the highest sensitivity in end-point titre. In sensitive pair optimization, it was observed that the mAb, b10c1, as a detector and the mAb, Aa3c10, as a capture antibody showed the highest absorbance to detect 50pg/ml PfHRP2 antigen. The prototype formulation of lateral flow assay using the mAbs (Aa3c10; b10c1) showed good reactivity with WHO panel and no false-positive results were observed with twenty clinically negative samples and five P. vivax positive samples. Conclusions: The novel monoclonal antibodies (Aa3c10, b10c1) against truncated PfHRP2, could be a strong potential candidates that can be included in making RDTs with better sensitivity and stability.
ABSTRACT
Background: Myocardial infarction which is an outcome measure in clinical trials, observational studies and quality assurance program have several conventional risk factors which include older age, hypertension, diabetes, decreased physical activity, alcohol intake, smoking, abdominal obesity, highrisk diet, psychological stress. Hypomagnesemia and means platelet volume is now recognized as a significant risk factor for atherogenesis, and thus for hypertension, ischemic heart disease, cardiac arrhythmias, coronary vasospasm, sudden cardiac death, cerebrovascular accident, and myocardial infarction. The aim of the study: To determining the relationship between serum Magnesium levels on platelet reactivity in Acute Myocardial infarction. Therefore in this study, we attempted to find the impact of serum Magnesium level on the Mean Platelet Volume and the use of these parameters as novel biomarkers to predict AMI. Materials and methods: A case-control study was carried out in the Department of Cardiology, Govt. Royapettah Hospital/ Kilpauk Medical College. Totally 88 Acute Myocardial Infarction patients (for the estimated prevalence of 30.36% in urban Indian population) admitted in the Intensive Coronary Care Unit between July to October 2015 and 88 age and sex-matched apparently normal individuals were included. Fasting venous peripheral blood samples were drawn within 48 hours of admission. Blood samples were taken into standardized trisodium citrate tubes (stored at room N. Jayaprakash, V. Madhavan, Aswanaa Kamanuru Govindarajulu. Serum magnesium levels and mean platelet volume (MPV) as biomarkers in acute myocardial infarction. IAIM, 2019; 6(4): 1-8. Page 2 temperature) and a sterile vacutainer which was serum separated, aliquoted into 2 Eppendorf’s and stored at minus 20 º C until further analysis. Results: Both the Systolic and Diastolic Blood pressure was significantly elevated in the cases (p= 0.000) which was statistically significant. The Fasting Blood Glucose levels were raised in AMI patients (p=0.001) although only 38.6% were known Diabetics. Urea levels were increased in cases (p=0.007) which was significant. The serum magnesium values were significantly lower in AMI patients in comparison to the normal individuals (p= 0.000) and the Mean Platelet Volume was significantly elevated in the cases than the control (p=0.004). Conclusion: Our study demonstrated that Magnesium levels were reduced in AMI patients and that Mean platelet Volume was elevated in AMI patients however a cause-effect relationship between the two parameters was not established. However, we propose that MPV and Magnesium may be useful biomarkers in identifying patients with increased risk for AMI. Further, a cohort study design including all the confounding variables can better address their relationship and role as adjuvant biomarkers in the diagnosis of AMI.