FILTRATION &
ULTRAFILTRATI
ON
FILTRATION
• Filtration is a physical or mechanical process
used to separate solids from liquids or gases by
passing the mixture through a filter medium that
allows only certain substances to pass through
while blocking others.
• The solid particles that do not pass through are
called the residue, while the liquid or gas that
passes through is called the filtrate.
FILTRATION IN BIOPHYSICAL SCIENCE
• In biophysical science, filtration is a crucial
technique used to separate biological
molecules, cells, and other microscopic
particles based on size, shape, or other
physical properties. It plays a key role in
research, medical applications, and industrial
biotechnology.
TYPES OF FILTRATION
1.Membrane Filtration
– Uses semi-permeable membranes with specific pore
sizes.
– Example: Ultrafiltration is used to concentrate
proteins or remove small contaminants from
biological samples.
2.Microfiltration (MF)
– Removes large particles like bacteria and cell debris.
– Example: Used in sterilization of culture media and
water purification.
3. Ultrafiltration (UF)
– Retains macromolecules such as proteins while allowing
smaller molecules like salts and water to pass.
– Example: Protein purification and separation in
biochemistry.
4. Nanofiltration (NF)
– Removes small organic molecules and some ions, often
used for water treatment.
– Example: Used in pharmaceutical processes to filter out
specific compounds.
5. Dialysis and Reverse Osmosis (RO)
– Uses selective membranes to remove toxins or
concentrate solutes.
– Example: Kidney dialysis removes waste from the
blood.
6. Centrifugal Filtration
– Uses centrifugal force and filtration to separate
substances based on density.
– Example: Separation of cell components in
laboratory research.
APPLICATIONS IN BIOPHYSICAL
SCIENCE:
1. Protein Purification: Isolating specific
proteins for structural and functional analysis.
2. Cell Culture: Filtering out bacteria and
viruses from media.
3. DNA and RNA Purification: Using
specialized filters to separate nucleic acids.
4. Medical Applications: Sterilization of fluids,
vaccines, and biopharmaceuticals.
5. Environmental Science: Removing
contaminants from air and water.
ULTRAFILTRATION
• Ultrafiltration is a membrane-based separation
technique widely used in biophysical and biochemical
research. It allows the selective separation of
molecules based on size, typically using semi-
permeable membranes with defined molecular weight
cut-offs (MWCO). This process is driven by pressure
or concentration gradients and is essential for protein
purification, buffer exchange, and macromolecule
concentration
• Principles of Ultrafiltration
• Membrane Selectivity: Ultrafiltration membranes
have specific pore sizes that allow smaller
molecules (e.g., salts, buffers, small metabolites) to
pass through while retaining larger molecules (e.g.,
proteins, nucleic acids, viruses).
• Driving Force: Uses pressure (centrifugal, gas, or
hydraulic) or osmotic gradients to facilitate
separation.
• Molecular Weight Cut-Off (MWCO): Defines the
smallest molecular weight (in Daltons) that is
retained (~90%) by the membrane. Common
MWCO values range from 1 kDa to 100 kDa.
Applications in Biophysical Science
• Protein Purification and Concentration
– Removes small molecules and salts while retaining
proteins of interest.
– Used for concentrating dilute protein solutions before
experiments like spectroscopy, electrophoresis, or
crystallization.
• Buffer Exchange and Desalting
– Replaces a protein's solvent environment without affecting
its structure or activity.
– Useful for preparing proteins for assays requiring specific
buffers.
• Separation of Biomolecules
– Differentiates molecules based on size, such as separating
proteins from nucleic acids.
– Employed in virus purification by filtering out host cell
debris.
• Cell-Free Extract Preparation
– Isolates cellular components while removing small
contaminants.
– Essential for proteomic and metabolomic studies.
• Lipid and Vesicle Research
– Used to concentrate or purify liposomes and exosomes.
– Ensures removal of free lipids from reconstituted
membrane systems
Techniques and Devices Used
• Centrifugal Ultrafiltration Devices: Small-
scale devices for rapid concentration and
buffer exchange.
• Tangential Flow Filtration (TFF): Large-
scale ultrafiltration used in bioprocessing.
• Pressure-Driven Filtration Systems: Used
for high-volume separations in industrial
applications.
Advantages and Limitations
• ✅ Advantages:
• Rapid and simple technique.
• No need for chemical additives.
• High recovery rates for macromolecules.
• ❌ Limitations:
• Membrane fouling can reduce efficiency.
• Some proteins may denature due to shear forces.
• Retention of some small molecules may not be
perfect.
• Ultrafiltration remains an essential tool in biophysical
science, enabling precise control over biomolecule
separation and preparation.
THANK YOU
