Molecular Biology Techniques: Blotting, Hybridoma, and Fermentation
Immunoblotting
Viral antigens are detected with a polyclonal or a monoclonal antibody (MAb) on nitrocellulose paper.
- After incubation, the protein bands (immune complexes) are visualized with peroxidase-conjugated protein and a color reagent.
- A color develops in the bands where the antibody binds to the antigen.
- Immunoblotting is an assay mixture of these two techniques.
Western Blotting
Western blotting is based on the principles of immunochromatography where proteins are separated in polyacrylamide
Read MoreMolecular Genetics and Systems Biology: Key Mechanisms
1. Molecular Mechanisms of Complex Genetic Diseases
Complex genetic diseases are multifactorial and result from the interaction of multiple genes and environmental factors. Key mechanisms include:
- Polygenic inheritance: Many genes contributing small effects.
- Genetic variants: SNPs and copy number variations altering gene function.
- Epistasis: Gene–gene interactions.
- Gene–environment interactions: Influence of diet or toxins.
- Epigenetic modifications: DNA methylation and histone changes regulating expression
Recombinant DNA Technology in Vaccine and Insulin Production
Recombinant DNA Technology in Vaccine Manufacture
Steps for Recombinant DNA Preparation
1. Isolation of Genetic Material or DNA
Genetic material is present inside the cells. It has to be obtained in pure form without even the attached histones and other proteins. To get the desired DNA, cells are treated with lysozyme (for bacteria), cellulase and pectinase (for plant cells), and chitinase (for fungal cells).
- The treated cells are homogenized and centrifuged to rupture the cells as well as nuclear envelopes.
DNA Restriction Digestion and Agarose Gel Electrophoresis
Understanding Restriction Enzymes
Restriction enzymes (RE), or restriction endonucleases, act as molecular scissors by cutting DNA at specific recognition sequences. These enzymes are isolated from various cellular strains and typically recognize palindromic sequences of 4 to 6 base pairs. Once the target site is located, the enzyme cleaves both strands of the DNA double helix, resulting in restriction fragments.
Common Restriction Enzymes
Every enzyme has unique target sites. For example, Lambda DNA
Read MoreDrug Physicochemical Properties and Biological Action
Physicochemical Properties in Relation to Biological Action
Physicochemical Properties refer to the physical and chemical characteristics of a drug molecule that influence its behavior in biological systems. These properties play a crucial role in determining the drug’s absorption, distribution, metabolism, and excretion (ADME). They include:
- Ionization
- Solubility
- Partition Coefficient
- Hydrogen Bonding
- Protein Binding
- Chelation
- Bioisosterism
- Optical and Geometrical Isomerism
1. Ionization
Ionization is a key
Read MorePharmacology Drug Classes and Mechanisms Summary
NSAIDs: Pain Relief and Inflammation Reduction
NSAIDs (Non-Steroidal Anti-Inflammatory Drugs) are a class of meds that reduce inflammation, pain, and fever. They’re like the ultimate pain relief squad.
Classification
- Non-selective COX inhibitors: Aspirin, Ibuprofen, Diclofenac, Naproxen
- COX-2 selective inhibitors: Celecoxib, Etoricoxib
Pharmacology of Aspirin
- Mechanism: Aspirin irreversibly inhibits COX-1 and COX-2 enzymes, reducing prostaglandin synthesis, which leads to decreased pain, inflammation,