Bioassay development
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Bioassay development is the process of obtaining a reliable assay to measure a substance's biological activity. It involves carefully studying the biological target, designing an appropriate test system, and understanding how the measurement relates to the substance's function. For example, an assay was developed to measure the activity of a growth factor (L) involving two test systems - a cell line expressing the receptor and an ELISA to detect receptor phosphorylation, which is proportional to the amount of active L. The final bioassay protocol is validated based on performance characteristics like accuracy, precision, sensitivity, and reproducibility.
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Reverse Pharmacognosy
This slide contains definition of pharmacognosy and reverse pharmacognosy, steps of pharmacognosy, parts of reverse pharmacognosy, comparison of pharmacognosy and reverse pharmacognosy, Selnergy, Selnergy in reverse pharmacognosy, application and advantages of reverse pharmacognosy, Selnergy on e-viniferin, Conclusion and Reference.
LEAD IDENTIFICATION BY SUHAS PATIL (S.K.)
This document provides an overview of lead identification in drug discovery. It discusses various methods for identifying lead compounds, including combinatorial chemistry, high-throughput screening, and in silico lead discovery techniques. Combinatorial chemistry allows for the rapid production and screening of large compound libraries. High-throughput screening assays test large numbers of compounds against biological targets using automated technologies. In silico methods like molecular docking use computer simulations to predict how compounds may bind and interact with targets. The goal is to find initial "hit" compounds that can then be optimized into drug candidates.
Biosensor libraries harness large classes of binding domains for construction...
The document summarizes research on developing novel transcriptional regulators (TFs) that can sense small molecules by fusing DNA-binding domains (DBDs) to ligand-binding domains (LBDs). The researchers generated a library of potential TFs by combining 15 DBDs with 15 LBDs using a ligase cycling reaction. They screened the library using flow cytometry to enrich for TFs that repressed GFP expression in the absence of inducer benzoate but allowed expression in its presence. Two top-performing TFs were characterized further and shown to repress promoter activity until benzoate was added, demonstrating their potential as biosensors.
Biological Assay .pdf
This document summarizes biological assays, specifically focusing on pharmaceutical quality management. It defines an assay as a procedure used to qualitatively or quantitatively assess the presence, amount, or functional activity of an analyte, such as a drug or biochemical substance. There are three main types of assays discussed: chemical assays, immunoassays, and bioassays. Bioassays are used to estimate the concentration or potency of a pharmaceutical drug by comparing its effects on animals or humans to a standard. The document outlines various techniques for bioassays including quantal, graded, and effect over time assays. It also discusses standards, accuracy, dependencies on dose and intensity, and applications of bioassays in pharmaceutical quality management.
Bioanalytical method development and validation .
1) A bioanalytical method was developed and validated for the quantification of levodopa and carbidopa in rat plasma using LC-MS/MS. Derivatization and ion-pairing chromatography were used to improve the chromatographic retention of the polar analytes.
2) The method was fully validated as per FDA guidelines and demonstrated selectivity, linearity, accuracy, precision, recovery, matrix effects and stability in accordance with acceptance criteria.
3) The validated method was successfully applied to support toxicokinetic studies of levodopa and carbidopa in rats.
Seminar on drug disposition
This document summarizes a seminar on computational methods for drug disposition. It discusses two approaches to modeling drug disposition: qualitative and quantitative. The quantitative approach uses pharmacophore modeling and docking to study drug interactions, while the qualitative approach uses QSAR and QSPR to correlate molecular descriptors with ADMET properties. The document also reviews the key processes of drug disposition: absorption, distribution, metabolism, and excretion. It provides examples of two research articles, one on the placental disposition of the immunosuppressant tacrolimus, and another on the pharmacokinetics of miltefosine in mice and hamsters infected with Leishmania.
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Biosensor libraries harness large classes of binding domains for construction...
Biosensor libraries harness large classes of binding domains for construction...
Bioassay development
- 1. BIOASSAY DEVELOPMENT BY PRAVESH VERMA M.PHARM[PHARMACOLOGY] 2nd SEM BABASAHEB BHIMRAO AMBEDKAR UNIVERSITY,LUCKNOW,UP,INDIA
- 5. to study a biological process,
- 6. to detect the presence and quantity of a substance in a sample,
- 7. and to screen for active molecules from a library of molecules.AIM
- 9. it has to be fully characterized.
- 10. The characterization of a substance requires the determination of its physiochemical properties by physiochemical assays (characterization)
- 12. A biological test system is required for a bioassayREQUIREMENT OF BIOASSAY
- 15. A variety of a bioassay can be developed for a given project
- 16. The most common bioassays are :-1.Biochemical assays with isolated proteins 2.Cell-based assays.
- 20. C.Understanding the selected measurement’s application range (boundary conditions) and making sure the intended uses are within the boundary D. Obtaining a control substance known to generate the intended responses in the test system E. Building algorithms that can mathematically relate the final detected signal to the intrinsic biological function
- 22. A growth factor (L) is a potential therapeutics found in drug discovery phase and it is currently under development.
- 23. It is known that L may denature in some formulation resulting in lose of activity
- 26. It is also known that L’s biological activity started with its binding to the cell surface receptor R.
- 27. The binding of L initiated the dimerization of receptor R on the cell surface.
- 28. This is followed by autophosphorylation of receptor R that is followed by many other biological responses
- 32. Here we choose to measure the phosphorylation of receptor R with a sandwiched enzyme-linked immunosorbent assay (ELISA) system.
- 33. In this assay, one antibody attached to the bottomsurface of a microplate captures solubilized receptor R.
- 35. The first test system is the cell line that expresses receptor R and is responsive to active L.
- 38. 9. Dynamic Range 10. Robustness 11. Boundary Conditions 12. Reproducibility 13. Scalability
Editor's Notes
- PRAVESH