This document discusses biological products and bioseparation techniques. It begins by defining different types of biologically derived products based on their chemical nature and applications. These include solvents, organic acids, vitamins, sugars, lipids, nucleic acids and various proteins. It then describes various cell disruption techniques used in bioseparation, including physical methods like bead mill, rotor-stator mill, French press, and chemical methods like detergent, enzyme, and solvent disruption. Finally, it discusses membrane-based bioseparation techniques like microfiltration, ultrafiltration, nanofiltration, and dialysis, explaining the separation mechanisms and operating parameters for each.
This lecture note describes the process of Effluent Treatment (ET). Emphasis is give to the biological aspects of ET. Free to reuse, remix, modify and share for non-commercial and commercial purposes.
Polyhydroxyalkanoates as an example of natural biodegredable polymers .
PHAs are biodegredable biopolyesters produced by a variety of gram negative and gram positive bacteria.
They have a variety of applications in the industrial and medical fields .
This lecture note describes the process of Effluent Treatment (ET). Emphasis is give to the biological aspects of ET. Free to reuse, remix, modify and share for non-commercial and commercial purposes.
Polyhydroxyalkanoates as an example of natural biodegredable polymers .
PHAs are biodegredable biopolyesters produced by a variety of gram negative and gram positive bacteria.
They have a variety of applications in the industrial and medical fields .
The following presentation is only for quick reference. I would advise you to read the theoretical aspects of the respective topic and then use this presentation for your last minute revision. I hope it helps you..!!
Mayur D. Chauhan
Process scale-up is a critical activity that enables a fermentation process achieved in research and development to operate at a commercially viable scale for manufacturing.
Microbial Kinetics in Batch Culture
Culture system containing a limited amount of nutrient, which is inoculated with the microorganism. Cells grow until some component is exhausted or until the environment changes so as to inhibit growth. Biomass concentration defined in terms of cell dry weight measurements (g/l) or total cell number (cells/ml).
Lineweaver-Burke Equation.....We remember the Monod Equation
Invert…
The equation now has the form of a straight line with intercept.
Y = MX + C
By plotting as a function of
You get a straight line, where the slope is , and the y–axis intercept is .
Product Yield Coefficient
Maintenance:
Cells use energy and raw materials for two functions, production of new cells and the maintenance of existing cells. In general, consumption of materials for maintenance is small w.r.t. the amount of materials used in the synthesis of new biomass.
Generally it is assumed that the use of materials for maintenance is proportional to the amount of cells present.
Overview
Industrial fermentations comprise both upstream (USP) and downstream processing
(DSP) stages. USP involves all factors and processes leading to and including the
fermentation. It consists of three main areas: the producer organism, the medium
and the fermentation process.
The following presentation is only for quick reference. I would advise you to read the theoretical aspects of the respective topic and then use this presentation for your last minute revision. I hope it helps you..!!
Mayur D. Chauhan
Process scale-up is a critical activity that enables a fermentation process achieved in research and development to operate at a commercially viable scale for manufacturing.
Microbial Kinetics in Batch Culture
Culture system containing a limited amount of nutrient, which is inoculated with the microorganism. Cells grow until some component is exhausted or until the environment changes so as to inhibit growth. Biomass concentration defined in terms of cell dry weight measurements (g/l) or total cell number (cells/ml).
Lineweaver-Burke Equation.....We remember the Monod Equation
Invert…
The equation now has the form of a straight line with intercept.
Y = MX + C
By plotting as a function of
You get a straight line, where the slope is , and the y–axis intercept is .
Product Yield Coefficient
Maintenance:
Cells use energy and raw materials for two functions, production of new cells and the maintenance of existing cells. In general, consumption of materials for maintenance is small w.r.t. the amount of materials used in the synthesis of new biomass.
Generally it is assumed that the use of materials for maintenance is proportional to the amount of cells present.
Overview
Industrial fermentations comprise both upstream (USP) and downstream processing
(DSP) stages. USP involves all factors and processes leading to and including the
fermentation. It consists of three main areas: the producer organism, the medium
and the fermentation process.
Recovery and purification of intracellular and extra cellular productsBangaluru
Product recovery and purification, such as centrifugal, chromatography, crystallization, dialysis, drying, electrophoresis, filtration, precipitation, etc., are essential finishing steps to any commercial fermentation process.
PHYTOSOMES AND ELECTROSOMES of novel drug delivery system .pptxnthanuja0331
Introduction and methods of preparation, evaluation parameters of phytosomes, advantages , disadvantages and applications of phytosome and electrosomes. some of marketed products of phytosomes.
This presentation explores a brief idea about the structural and functional attributes of nucleotides, the structure and function of genetic materials along with the impact of UV rays and pH upon them.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
Future Directions: Continued research and technological advancements hold promise for new treatments and applications.
Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
Cancer cell metabolism: special Reference to Lactate PathwayAADYARAJPANDEY1
Normal Cell Metabolism:
Cellular respiration describes the series of steps that cells use to break down sugar and other chemicals to get the energy we need to function.
Energy is stored in the bonds of glucose and when glucose is broken down, much of that energy is released.
Cell utilize energy in the form of ATP.
The first step of respiration is called glycolysis. In a series of steps, glycolysis breaks glucose into two smaller molecules - a chemical called pyruvate. A small amount of ATP is formed during this process.
Most healthy cells continue the breakdown in a second process, called the Kreb's cycle. The Kreb's cycle allows cells to “burn” the pyruvates made in glycolysis to get more ATP.
The last step in the breakdown of glucose is called oxidative phosphorylation (Ox-Phos).
It takes place in specialized cell structures called mitochondria. This process produces a large amount of ATP. Importantly, cells need oxygen to complete oxidative phosphorylation.
If a cell completes only glycolysis, only 2 molecules of ATP are made per glucose. However, if the cell completes the entire respiration process (glycolysis - Kreb's - oxidative phosphorylation), about 36 molecules of ATP are created, giving it much more energy to use.
IN CANCER CELL:
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
Unlike healthy cells that "burn" the entire molecule of sugar to capture a large amount of energy as ATP, cancer cells are wasteful.
Cancer cells only partially break down sugar molecules. They overuse the first step of respiration, glycolysis. They frequently do not complete the second step, oxidative phosphorylation.
This results in only 2 molecules of ATP per each glucose molecule instead of the 36 or so ATPs healthy cells gain. As a result, cancer cells need to use a lot more sugar molecules to get enough energy to survive.
introduction to WARBERG PHENOMENA:
WARBURG EFFECT Usually, cancer cells are highly glycolytic (glucose addiction) and take up more glucose than do normal cells from outside.
Otto Heinrich Warburg (; 8 October 1883 – 1 August 1970) In 1931 was awarded the Nobel Prize in Physiology for his "discovery of the nature and mode of action of the respiratory enzyme.
WARNBURG EFFECT : cancer cells under aerobic (well-oxygenated) conditions to metabolize glucose to lactate (aerobic glycolysis) is known as the Warburg effect. Warburg made the observation that tumor slices consume glucose and secrete lactate at a higher rate than normal tissues.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
Introduction:
RNA interference (RNAi) or Post-Transcriptional Gene Silencing (PTGS) is an important biological process for modulating eukaryotic gene expression.
It is highly conserved process of posttranscriptional gene silencing by which double stranded RNA (dsRNA) causes sequence-specific degradation of mRNA sequences.
dsRNA-induced gene silencing (RNAi) is reported in a wide range of eukaryotes ranging from worms, insects, mammals and plants.
This process mediates resistance to both endogenous parasitic and exogenous pathogenic nucleic acids, and regulates the expression of protein-coding genes.
What are small ncRNAs?
micro RNA (miRNA)
short interfering RNA (siRNA)
Properties of small non-coding RNA:
Involved in silencing mRNA transcripts.
Called “small” because they are usually only about 21-24 nucleotides long.
Synthesized by first cutting up longer precursor sequences (like the 61nt one that Lee discovered).
Silence an mRNA by base pairing with some sequence on the mRNA.
Discovery of siRNA?
The first small RNA:
In 1993 Rosalind Lee (Victor Ambros lab) was studying a non- coding gene in C. elegans, lin-4, that was involved in silencing of another gene, lin-14, at the appropriate time in the
development of the worm C. elegans.
Two small transcripts of lin-4 (22nt and 61nt) were found to be complementary to a sequence in the 3' UTR of lin-14.
Because lin-4 encoded no protein, she deduced that it must be these transcripts that are causing the silencing by RNA-RNA interactions.
Types of RNAi ( non coding RNA)
MiRNA
Length (23-25 nt)
Trans acting
Binds with target MRNA in mismatch
Translation inhibition
Si RNA
Length 21 nt.
Cis acting
Bind with target Mrna in perfect complementary sequence
Piwi-RNA
Length ; 25 to 36 nt.
Expressed in Germ Cells
Regulates trnasposomes activity
MECHANISM OF RNAI:
First the double-stranded RNA teams up with a protein complex named Dicer, which cuts the long RNA into short pieces.
Then another protein complex called RISC (RNA-induced silencing complex) discards one of the two RNA strands.
The RISC-docked, single-stranded RNA then pairs with the homologous mRNA and destroys it.
THE RISC COMPLEX:
RISC is large(>500kD) RNA multi- protein Binding complex which triggers MRNA degradation in response to MRNA
Unwinding of double stranded Si RNA by ATP independent Helicase
Active component of RISC is Ago proteins( ENDONUCLEASE) which cleave target MRNA.
DICER: endonuclease (RNase Family III)
Argonaute: Central Component of the RNA-Induced Silencing Complex (RISC)
One strand of the dsRNA produced by Dicer is retained in the RISC complex in association with Argonaute
ARGONAUTE PROTEIN :
1.PAZ(PIWI/Argonaute/ Zwille)- Recognition of target MRNA
2.PIWI (p-element induced wimpy Testis)- breaks Phosphodiester bond of mRNA.)RNAse H activity.
MiRNA:
The Double-stranded RNAs are naturally produced in eukaryotic cells during development, and they have a key role in regulating gene expression .
Earliest Galaxies in the JADES Origins Field: Luminosity Function and Cosmic ...Sérgio Sacani
We characterize the earliest galaxy population in the JADES Origins Field (JOF), the deepest
imaging field observed with JWST. We make use of the ancillary Hubble optical images (5 filters
spanning 0.4−0.9µm) and novel JWST images with 14 filters spanning 0.8−5µm, including 7 mediumband filters, and reaching total exposure times of up to 46 hours per filter. We combine all our data
at > 2.3µm to construct an ultradeep image, reaching as deep as ≈ 31.4 AB mag in the stack and
30.3-31.0 AB mag (5σ, r = 0.1” circular aperture) in individual filters. We measure photometric
redshifts and use robust selection criteria to identify a sample of eight galaxy candidates at redshifts
z = 11.5 − 15. These objects show compact half-light radii of R1/2 ∼ 50 − 200pc, stellar masses of
M⋆ ∼ 107−108M⊙, and star-formation rates of SFR ∼ 0.1−1 M⊙ yr−1
. Our search finds no candidates
at 15 < z < 20, placing upper limits at these redshifts. We develop a forward modeling approach to
infer the properties of the evolving luminosity function without binning in redshift or luminosity that
marginalizes over the photometric redshift uncertainty of our candidate galaxies and incorporates the
impact of non-detections. We find a z = 12 luminosity function in good agreement with prior results,
and that the luminosity function normalization and UV luminosity density decline by a factor of ∼ 2.5
from z = 12 to z = 14. We discuss the possible implications of our results in the context of theoretical
models for evolution of the dark matter halo mass function.
6. 1. Separation on chemical processes.
2. Low molecular weight compound such as amino acid , vitamins are purified
by conventional processes. These are:
liquid-liquid extraction,
packed bed adsorption,
evaporation
drying with practically no necessary modificationss.
NATURE OF BIO SEPARATION
7. Bases of separtion would be.,
Density, size, porosity, shape , polarity, solubility, diffusivity, volatality,
charges, molecular weight, partition coeeficent,l ight absorption
DIFFERENCES BETWEEN BIOLOGICAL SEPARATION AND
SYNTHETIC SEPARTTION
I. Large volume of dilute streams are needed even for low
concentration compound separation.,ex : For monoclonal
antibodies in 0.1mg/ml in mammalian cell culture supernatants.
II. Bioseparation should be Selective in nature due to similiarty
between impurities and product of interest.
III. Potential to avoid extreme physiochemical conditions., pH,
temperature etc due to degradtion of biological products.
IV. Sometimes sub – ambient temperature required for heat labile.
V. Solutions are prepared inside bioprocess for inectables
therapeutics and endotoxin free compound
8. CELL DISRUPTION
The plasma membrane of different oragnism can be destroyed or disrupted by
using solvents,detergents,osmoticshock etc.It can be physical or chemical.
Disrupted cells can be gram negative, gram positive, mould cells,yeast
cells,mammalian cells,cultured plant cells and ground cells etc.
The main barrier is the cell wall which is composed of peptidoglycan,
teichoic acid and polysaccharides and is about 0.02 to 0.04 microns thick.
• Gram negative have multilayered envelops and thinner peptodoglycan.
• Yeasts which are unicellular have thick cell walls, typically 0.1 to
0.2 microns in thickness (such as glucans, mannans and chitins.)
They are composed of phospholipids arranged in the form of a bi-layer with the
hydrophilic groups of the phospholipids molecules facing outside .
The hydrophobic residues remain inside the cell membrane where they are
shielded from the aqueous environment present both within and outside
the cell.
9.
10. PHYSICAL METHOD
Disruption in bead mill
Disruption using a
rotor-stator mill
Disruption using French
press
Disruption using
ultrasonic vibrations
Disruption using
detergents
Disruption using
enzymes e.g. lysozyme
Disruption using
solvents
Disruption using
osmotic shock
CHEMICAL METHOD
13. BEAD MILL METHOD
This device consists of a stationary
block with a tapered cavity stator and
a truncated cone shaped rotating object
called the rotor.
Typical rotation speeds are in the
10,000 to 50,000 rpm range.
The cell suspension is fed into the tiny
gap between the rotating rotor and the
fixed stator.
High rate shear generated in the space
between the rotor and the stator as
well as the turbulence thus generated
are responsible for cell disruption.
ROTOR-STATOR MILL
14. FRENCH PRESS METHOD
It consist of cylinder fitted
with plunger connencted
to hydraulic press.
Cells placed inside cylinder
and pressurized (10k to 50k
psi) using plunger ,
through an orfice
suspension cells emerges at
high velocity due to
primary shear stress cell
starts disrupting.
Ultrasonic vibrations of
frequency greater than
18kHz are used to disrupt
cells.
Vibrations create
cavitation i.e., formation of
bubbles and they reach
resonance size where they
collapse releasing
mechanical energy in form
of shock, and disrupts the
cell membrane.
ULTRASONIC VIBRATION
15. DETERGENT DISRUPTION
They disrupts the cell
membranes and phosholipids
of mammalian cells mainly.
Bacterial celle needs
conjunction with lysozyme to
disrupts by weaken its walls.
Non ionic for bioprocessing
havinf least damage effect
such as tween series
detergents.
Need for removal.
Enzymes are used to
destroy the cell membrane.
Such as lysozymes,
pectinase used for
disruption and breaking
the bonds between the
membranes.
But they are costly.
Need for removal after
action.
ENZYMES DISRUPTION
16. OSMOTIC SHOCK
Semi premeable cell
membrane are transferred
into hypotonic solution
from isotonic that results
into rapid expansion of
cell & ruptures called
osmotic shock.
Used to lyse mammalian
cells,remove periplasmic
substances by expelation.
Acetone act on cell
membrane solubilizing
its phosholipids and
denature the protein.
Toluene for fungalcells.
Important to remove by
volatility.
ORGANIC SOLVENT
17. Precipitation based bioseparation essentially involves selcective conversion of a specific
dissolved component of a complex mixture to an insoluble form using appropriate physical
or physicochemical means.
The insoluble form which is obtained as a precipitate is sepatated from the dissolved
components by appropriate solid-liquid separation techniques such as centrifugation.
Biological macromolecules can be precipitated by:
1. Cooling
2. pH adjustment
3. Addition of solvents such as acetone and ethanol
4. Addition of anti-chaotropic salts such as ammonium
sulphate and sodium sulfate
5. Addition of chaotropic salts such as urea
and guanidine hydrochloride
6. Addition of biospecific reagents as in immunoprecipitation
PRECIPITATION
18. USING ORGANIC
SOLVENTS
• By reducing dielectric constant
USING ANTI-CHAOTOPIC
SALTS
• By decreasing solubility of proteins
PRECIPITATION METHODS
A.
B
.
19. Solvent based precipitation method
reduced the dielctric constant of
medium in which they are present.
ln {S/Sw}={[A/RT ][1/ew]- [1/e]}
S= solubility of protein, Sw =
solubility of protein in water, A=
constant, e = dielectric constant of
medium, ew = dielectric constant of
water.
Protein have lower solubiltiy in
medium[ethanol] than in water.
lower concentration of organic
solvent are used in precipitation
processes at low temperature
Ex; human plasma protein
purification by cohn fractionation.
Reducing protein solubility by
increasing salt concentration results
in increase in protein protein
hydrophobic interaction.
ln(S)=B – Ks Cs i.e., Cohnequation
B= Constant, Ks= salting out
constant, Cs= salt concentration
At low temp. 4degree celsius, &
constant depend on salt, pH and
protein solution.
More protein precipiated out &
stability increases at low temp. by
synergetic rxn.
Salt used = Ammonium salts, NaCl
Can be dirct addition or saturated
Ex; 30% -50% Ammoniumsalt cut.
A. B.
20. STAGES OF PRECIPITATION:
1. Mixing
2. Nucleation
3. Diffusion limited growth
4. Convection limited growth
• Formation of homogenous mixture and mix it .Time needed for mixiing can
be determined by: Tm= (l)(l)/4D ; l= avg eddy length, D= diffusivity.
• At supernaturation minute particles form called nucleation .
• Diffusion limited growth increases collision between particel hence increses
rate of fomation of bigger particles in microns.
• Mixing directly proportional to frequency of collision.
• Aging process takes place i.e., time given to precipitate & supernatant
formation( 6h-12hrs at 4 degree celsius)
Mechanism
22. Membranes are divided into:
1. Symmetric- similar compostition and morphology
2. 2.Asymmetric- non-identical composition
Basically membranes are:
1. Flat sheet membrane
2. Tubular membrane
3. Hollow fibre membrane
23. 1. MICROFILTRATION
This method is used for separation of fine particles from
solutions. The transmembrane pressure ranges from 1 to 50 psig. Most
microfiltration membranes capture particles by surface filtration, i.e. on
the surface of the membrane. In some cases depth filtration is also used.
Uses: for clarification, sterilization and slurry concentration.
2. ULTRAFILTRATION
Membranes retain macromolecules such as proteins while allowing
smaller molecules to pass through.
(a) separate large molecules from solvents,
(b) separate large molecules from smaller molecules,
(c) separate large molecules from one another.
The primary separation mechanism is size exclusion, but
physicochemical interactions between the solutes and the membrane, and
operating conditions can influence the process quite significantly.
Pressure ranges from 10 to 100 psig.
Most UF membranes are asymmetric.
24. 3. NANOFILTRATION
Allow salts and other small molecules To pass through but retain
larger molecules such as peptides, hormones and sugars.
Pressure in NF ranges from 40 to 200 psig.
MostNF membranes are composite i.e. asymmetric.
4.Liquid membrane processes
TRansport of solutes across a thin layer of a third liquid interposed
between two miscible liquids.
liquid membranes:
1. Emulsion liquid membranes (ELM)
2. Supported liquid membranes (SLM)
25. 5. MEMBRANE CHROMATOGRAPHY
Adsorption and chromatographic separations are
traditionally carried out using packed beds.
Separation mechanisms These include; 1.
Affinity binding 2. Ion-exchange interaction
3.Hydrophoboic
26. Diagram showing dialysis
Solute separation occurs
primarily because
smaller solutes partition
into the membrane
better than bigger
solutes because the
degree to which the
membrane restricts the
entry of solutes into it
increases with solute
size.
Smaller solutes also
diffuse more rapidly than
larger ones.
6.DIALYSIS
27. REFERENCES:
Research gate
NCBI
WIKIPEDIA
BOOK- PRINCIPLES OF BIO- SEPARATION ENGINEERING BY
RAJA GHOSH
BIOCHEMISTRY TECHNIQUES BOOK BY STRYER
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