Biotechnology Practical Course Overview

Biotechnology - Practical Course

• Biotechnology is the application of biological systems, living organisms, or derivatives to make or modify products for specific use.
• Medical biotechnology focuses on improving human health using living cells to produce pharmaceutical drugs or combat diseases.
• Medical biotech includes using DNA to manipulate genetic makeup for increased production of helpful human cell products (e.g., insulin).
• Agricultural biotechnology improves crops, increasing yields or introducing characteristics to withstand stress factors (weather, pests).
• Industrial biotechnology uses microorganisms or biological substances (e.g., enzymes) for industrial processes (paper, chemicals, textiles).
• Environmental biotechnology uses microorganisms to prevent, treat, and monitor environmental pollution, and remediate waste through biotreatment.
• Bioremediation is the use of microorganisms to clean up soil and groundwater.
• Growth Curve stages include lag phase, exponential phase, stationary phase, and decline phase.
• Colony-forming units (CFUs) are used for counting bacteria via counting colonies on a plate after dilution.
• Optical density measures absorbance of light passing through a culture, used to estimate bacterial cell count.
• Pure cultures are maintained and preserved through methods like subculturing, refrigeration, and cryopreservation.

Principal Steps of Bioengineered Biotechnology Process

• Upstream processing involves preparing microorganisms and raw materials needed for the process.
• Fermentation and transformation is the process of using a larger bioreactor to grow microorganisms that will create a desired product (e.g., antibiotic, protein).
• Downstream processing involves recovery and purification of the desired product from the medium or cell mass.
• Choosing organisms for industrial use involves selecting microorganism(s) with high growth rate, stability, high yield, and non-toxicity to produce desired product.

Methods for Selection and Isolation of Microorganisms

• Chemical methods include enrichment culture, use of minimal nutrients, or inhibitors (dyes, salts, metals, antibiotics).
• Physical methods include heating to kill vegetative cells (80°C), incubation at different temperatures (for thermophiles, mesophiles, psychrophiles) and observing pH preferences to isolate different groups of microorganisms.
• Biological methods include using different organisms and specific techniques for cultivating microorganisms.

Screening of Microorganisms

• Screening techniques involve isolating and identifying microorganisms from mixed populations.
• Examples of techniques: using natural sources (soil, milk), screening for organic acid/amine or antibiotic producers.
• Techniques might include dilution series and spreading on agar plates.

Biotechnological Process- Stages

• Stage 1 (upstream processing) involves preparing specific liquid cultures, sterilizing, and purifying materials.
• Stage 2 (fermentation) is growing microorganisms using bioreactors or fermenters (liquid or solid medium) to create desired products.
• Stage 3 (downstream processing) is isolating and purifying the desired product (antibiotics, proteins, etc)
• Fermenters (bioreactors) are vessels containing conditions for optimum growth and production (pH, temp, oxygen) and are typically either submerged or surface (solid-medium) cultures.

Fermenter/Bioreactor Components

• Typical fermenters have four or more baffles, devices for temperature control and aeration.
• Mechanical agitators are typically used for mixing, aeration, and distributing nutrients, and oxygen supply.
• Components include: inoculation pipe, air opening, stirring apparatus, pH probes, and temperature probes.

Fermentation Product Extraction

• Extraction considers factors such as final product value, required purity and properties of impurities to determine downstream processing stages.
• Stages include removal of insoluble components(solids), product isolation (separation, filtration, coagulation, etc - using different types of filtration), product purification (chromatography, crystallization, filtration), and product polishing (e.g. drying, sterilizing).

Molecular Biotechnology

• Molecular biotechnology involves transferring genetic information between organisms.
• Recombinant DNA technology is used to create new products or improved commercial processes through genetically modified organisms.
• Molecular biotechnology has applications in producing drugs, vaccines, diagnostics (e.g. diagnostics for infectious diseases), crops resistant to pathogens, and livestock with enhanced traits.

Recombinant DNA Technology

• Recombinant DNA technology, also called gene cloning, moves genetic information from one organism to another
• Cloning vector is essential to allow the gene construct to be amplified.
• Transforming host cells to take up the DNA construct and produce the required product (e.g protein)
• Techniques include DNA isolation and purification.

PCR(Polymerase Chain Reaction)

• PCR is a technique for amplifying DNA sequences.
• Steps include Denaturation, annealing and elongation to amplify specific DNA regions.

Molecular Diagnostics

• Molecular diagnostics uses methods for detecting and quantifying proteins, antibodies, and hormones.
• Examples include ELISA (enzyme-linked immunosorbent assay) techniques.
• Techniques can be used to identify and quantify genetic information to aid in pathogen detection, disease identification, and determining drug response.

Gene Therapy

• Gene therapy utilizes nucleic acids for treating diseases.
• Several strategies are used for gene transfer directly into the cytoplasm/nucleus (viral/nonviral delivery systems).
• Gene therapy can be classified as in vivo or ex vivo depending on whether genetic modification occurs within the body or in a cell culture.

Real Time PCR

• Real-time PCR is a quantitative method used to measure amplified DNA.
• Different detection methods exist, like SYBR Green or TaqMan probes.
• Multiplex PCR allows simultaneous detection of multiple DNA targets.

Immunological Diagnostic Procedures

• ELISA is a plate based assay for measuring soluble substances.
• Types include direct, indirect, sandwich, and competitive ELISA, each with specific design and application.
• Ouchterlony test is an immunodiffusion assay used for antibody and antigen detection..

Bioinformatics

• Bioinformatics combines biology, computer science, and information technology to study large biological data sets.
• Bioinformatics applications include gene detection, genomics, drug discovery, and disease prediction.

This quiz explores various fields within biotechnology, including medical, agricultural, industrial, and environmental applications. You'll learn about techniques such as genetic manipulation and bioremediation, which are crucial in advancing health and sustainability. Test your knowledge on how biotechnology impacts various sectors and the science behind it.