The document discusses nanofiber technology for protein purification using electrospun nanofibers and simulated moving bed chromatography. Specifically, it notes that electrospun nanofibers can produce high surface area materials allowing for rapid convective flow and high binding capacities. When packed into a filter and used with simulated moving bed chromatography, it allows for continuous high throughput protein purification at lower costs compared to traditional bead-based methods. The technology has potential for scaling up antibody purification for the biopharmaceutical industry.

1. Puridify
Nanofibre Technology
Christian Ackah
Patcha Chainiwatana
Kuan-Lin Chiang
Shreya Shah
Phillippa Shmeis
Shuiab Zariat

2. What Is So Important About This Technology?
• 2 technologies are Electrospun Nanotechnology (Cellulose Nanofibers) & Simulated Moving Bed(SMB)
• Electrospun polymeric fibers with sub micron diameters, produce remarkably high surfaces areas
permitting rapid convective flow operations and higher binding capacities.
• Appropriate packing of these nanofibers has allowed reproducible bind-elute operations at flow rates of
2,400 cm/h
• Thus High throughput and high binding capacity leads us down the path of continuous processing
• SMB system has proven itself in other industries e.g Chemical industry and operates very well in
continuous mode.
• SMB works on the basis of counter currents between stationary phase and mobile phase greatly enhancing
separation making the process more efficient
• Most importantly this system reduces costs
Why Are We Pushing For This Technology?
• More Involved in the fastest growing sector within the world of pharmaceuticals(biotechnology)
• 20% of market shares $153bn(2012) Set to grow to 41% by 2018 $215bn (Strickland, 2012)
• 200 monoclonal antibodies on the market, over 1000 in clinical trials (Rader, 2013)
• Antibody purification systems will very soon be in high demand
• Growth in fermentation titre 0.5gL to 50gL (Lightfoot and Moscariello, 2004 and Roque et al., 2004))
• Downstream has not yet matched upstream advancements in antibody purification
• The key to advancing downstream processing is said to be heavily reliable on bind and elute
chromatography unit operations, in economical terms. (Labrou and Clonis, 1994 and Lowe, 2001)
Christian Ackah

3. Benefits of Nanofibre Technology (FibroSelect)
● Reduces the running time of the process and hence saves on
manufacturing costs.
● Convective mass transfer - increased throughput - higher
productivity - more product collected over a short period of time.
● Saves money on materials, requires less reagent due to the
efficiency and lifespan of the nanofibres (fouling resistant).
Remains clean and is reusable.
● Potential to move away from batch separation towards continuous
downstream processing due to the high flow rate, combined with
SMB we could use a type of moving belt system to continuously
load a feed stream while recovering product simultaneously.
● FibroSelect purification platform is ideally suited to support the
industry’s increasing interest in single use and for smaller, more
flexible manufacturing facilities that can produce multiple products.
● 3D nanofibre compared to highly porous beads are 10 times more
efficient, and could reduce production costs by 25%.
● Allows for a more robust & efficient process – This will enable
drugs previously not developed due to cost to reach mass
manufacture.
Shuiab Zariat
Packing of nanofibre into a filter holder system
ready for bioseparation
Grad.ucl.ac.uk,.'ResearchImagesAsArt2012Gallery-UCLGraduateSchool'
Gov.uk,. 'Success Story: Puridify Heralds New Generation Of Biotherapeutics - Case Study - GOV.UK'. N.p., 2015. Web. 13 Dec. 2015.

4. comparison between past(beads) and future
(nanotechnology)- value offering
beads
structure and function relationship: diameter (10-100
micrometer) /diffusion limited→ slower flow
rate/lower productivity.
structure fouling relationship: resins which may
degrade to surface coating and channels in porous
cast membranes→ higher fouling chances.
structure lifespan relationship: resins usually age with
re-usage,(oxidants,hydraulic shocks,chlorine and
chloramine degradation.osmotic shocks,backwash)
all these will decrease the lifespan of resins .
narrow range of material : most commercial resins are
made of polystyrene sulfonate, or protein A from
staphylococcus aureus,...thus beads are more
expensive especially with the fact of producing
them.
nanofiber
structure and function relationship:
diameter(<1 micrometer however according to NSF
nanofiber have at least one dimension of 100
nanometer) / high surface area/ convective mass
transfer→ faster flow rate/ higher productivity.
structure fouling relationship: since nanofiber are
highly open porous nature→ reduce chances of
fouling.
structure lifespan relationship: due to its high
adsorbents properties the nanofiber remain clean
through resusage,longer life span .
wide range of material: nylon,acrylic polycarbonate,
polysulfonate, fluoropolymer,..... nanofibers are
produced in a cost effective manner especially with
the new techniques for their production
lower pressure drop.
Hardick, Oliver et al. 'Nanofiber Adsorbents For High Productivity Continuous Downstream Processing'. Journal of Biotechnology 213 (2015): 74-82. Phillippa jamal el din Shmeis

5. Nanofibre fabrication in a temperature and humidity
controlled environment for improved fibre consistency
● Electrospun cellulose acetate nanofibers
● Temperature range: 17-35o
C; Humidity range: 20-70% relative humidity
○ 9 conditions over 3 separate days
● Differential scanning calorimetry → evaluate thermal properties & melt enthalpy
● Scanning electron microscopy → images → average fibre diameter → porosity
● Controlled atmospheric conditions affect fiber quality
● Best quality of fibre produced at 25o
C, 50% RH:
○ Diameter uniformity, low beading, low diameter, increased surface area & pore size
● Reproducibility
○ Consistent observations in all 3 days
○ Way to control fibre morphology
○ Critical parameters: humidity, polymer solution
○ Validation, regulation, product specification
○ Enable mass production for commercial purposes
Patcha ChainiwatanaHardick, O; Stevens, B; Bracewell, DG; (2011) Nanofibre fabrication in a temperature and humidity controlled environment for improved fibre consistency. J MATER SCI , 46 (11) 3890 - 3898

6. (Diethylaminoethyl)
Simulated Moving Bed (SMB) system

8. challenges facing nanofiber technology
CHALLENGES IN NANOFIBERS
The vapors emitting from electrospinning solution while forming the web need to be recovered or
disposed of in an environmental friendly manner. This involves additional equipment and cost.
The fineness of fiber and evaporated vapor also raises much concern over possible health hazard
due to inhalation of fibers.
Thus the challenges faced can be summarized as:
● Economics
● Health hazards
● Solvent vapor
● Packaging shipping handling
Phillippa jamal el din ShmeisHegde,R.,Dahiya,A., Kamath,M.,Nanofiber Nonwoven,June 13,2005

9. ● Protein purification: affinity membrane chromatography
○ Electrospun nonwoven nanofibre membrane,
with ligands attached on surface
○ Regenerated cellulose nanofibre membrane
■ Hydrophilic, low nonspecific protein adsorption
■ High water permeability, large surface area,
■ Flow distribution, fouling resistance
■ Allow most proteins to covalently bind
● Potentially feasible to scale up the continuous process
○ Increase productivity using frequent,
short bind-elute cycles
○ Simulated moving bed (SMB)
In essence,
Patcha Chainiwatana
Ma Z and Ramakrishna S (2008). Electrospun regenerated cellulose nanofibre affinity membrane functionalised with protein A/G for IgG purification, J Membrane Sci. 319, 23-8.
Hardick, Oliver et al. 'Nanofiber Adsorbents For High Productivity Continuous Downstream Processing'. Journal of Biotechnology 213 (2015): 74-82.

10. ● Grad.ucl.ac.uk,. 'Research Images As Art 2012 Gallery - UCL Graduate School'. N.p., 2015. Web. 13 Dec. 2015.
● Gov.uk,. 'Success Story: Puridify Heralds New Generation Of Biotherapeutics - Case Study - GOV.UK'. N.p., 2015. Web. 13 Dec. 2015.
● Hardick, Oliver et al. 'Nanofiber Adsorbents For High Productivity Continuous Downstream Processing'. Journal of Biotechnology 213
(2015): 74-82.
● Hardick, O; Stevens, B; Bracewell, DG; (2011) Nanofibre fabrication in a temperature and humidity controlled environment for improved fibre
consistency. J MATER SCI , 46 (11) 3890 - 3898
● Hegde,R.,Dahiya,A., Kamath,M.,Nanofiber Nonwoven,June 13,2005
● Ma Z and Ramakrishna S (2008). Electrospun regenerated cellulose nanofibre affinity membrane functionalised with protein A/G for IgG
purification, J Membrane Sci. 319, 23-8.
● Mirko Faccini, Guadalupe Borja, Marcel Boerrigter, et al., “Electrospun Carbon Nanofiber Membranes for Filtration of Nanoparticles from
Water,” Journal of Nanomaterials, vol. 2015, Article ID 247471, 9 pages, 2015. doi:10.1155/2015/247471
● Nanoscience.com, (2015). Nanofiber Research for Nonwoven Textiles. [online]
References