fyp @ beans lab info session · the beans lab. bioengineered and applied nanomaterials laboratory....
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FYP @ BeANs LabInfo Session
2011 – 2012
The BeANs LabBioengineered and Applied Nanomaterials Laboratory
Logistics• Lab: N1.3-B3-15
• The mentors– 1 Postdoc– 3 Graduate Students
• The supervisor– Asst. Prof. Sierin Lim– N1.3-B3-06– +65 +514 1085– slim@ntu.edu.sg
• Website: www.ntu.edu.sg/home/slim
Projects
Future Healthcare• Drug delivery
system• MRI contrast
agent
Sustainable Earth• Photovoltaic
Cells
PROTEIN
The protein cages & chlorophasts
Image: Dalmau & Lim, et al., Biotechnology & Bioengineering,
vol. 104(4), 654-664 (2008)
Image: Kichoefer et al., PNAS, vol.102(12), 4348-52 (2005)
Image: Johnson et al., Structure, vol. 13, 637–648 (2005)
Image: Wikipedia
Chloroplast2-10 x 1 µm
Vault40 x 70 nm96 subunits
E225 nm
60 subunits
Ferritin15 nm
24 subunits
Project Organizations
DNAMolecular cloning(PCR, mutagenesis)
Expression / ProductionFermentation
(Growth condition optimization)
PurificationFPLC
(IEX, HIC, SEC, ultrafiltration)
CharacterizationStructure
(TEM, DLS, CD)Activity
(Colorimetric assay)
ModificationFunctionalization(ligand attachment)
Further AnalysisSequence comparison
(data mining, phylogenetic tree)
Future HealthcareScreening
High efficiency photosystem
Isolation
PrototypeDesign
ImmobilizationCurrent measurement
Sustainable Earth
BIE/11/067: Specific binding of anticancer drug with a carrier protein
• Ferritin – Iron storage protein– Converts metallic (Fe/Mn) salt to
magnetic nanoparticle that is a potential MRI contrast agent.
– Nonspecific encapsulation of limited number of drug molecules is possible in vitro
• Objectives:– Engineering of metal
binding peptide to the ferritin.
– Specific binding of metal-based anticancer agents
– Study of anticancer activity
Simsek et al. J Magn. Magnetic Mater. 293: 509 (2005)Sana et al. Biointerphases 5:FA48-FA52 (2010)
• Vault – natural nanocapsule for drug delivery– Composed of major vault protein (MVP, >70%), vault poly
(ADP-ribose) polymerase (VPARP) - Includes minimal interaction domain (mINT domain), telomerase-associated protein 1 (TEP1), untranslated RNA
– MVP (blue) domain 3 to 5 interacts specifically with mINT(golden). The region is referred to as iMVP.
– Controlling the dissociation of mINT has an implication in controlling the release of therapeutic agents.
BIE/11/068: Engineering MVP-mINT protein-protein interaction
• Objectives:– Identification of residues of the
protein-protein interaction sites– Modifying the interaction by site
directed mutagenesis– Comparing the mutant and wild
type interaction at specific pH
Kickhoefer, V.A., et al., PNAS,
2005. 102(12): p. 4348-4352.
BIE/11/069: Investigation of E2 protein surface charge to facilitate drug delivery
pH change
or
Salt ions
Zeta Potential ?
• E2 protein cage: Potential application as drug capsule
via cell endocytosis The change of zeta potential make E2
cage more attractive to target cell pH adjustment or varying ions cause
the change of zeta potential on E2 surface
• Objectives:– To investigate the surface charge
of E2 protein in different pH and pH of buffers by measuring its zeta potential
– Modification of the surface charge by substituting amino acids
– In vitro investigation on the E2 with most attractive feature to cancer cell
• Objectives:– Increase photo
electrochemical cell performance.
– Investigation on chloroplast and algae stability in ionic liquid.
• Photosynthesis– Electron production based on water
splitting reaction in the presence of light.
– Low stability of oxygen evolving catalyst in the presence of high ion (Na+, K+, Cl-) concentration.
– Ionic liquid as a substitution of ionic solution
BIE/11/070: Implication of ionic liquid as anolyte
Alberts, B., et al., Molecular Cell Biology of the Cell (4th Edition). 2002: Garland ScienceLam, K.B., et al., Sensors and Actuators, B: Chemical, 2006. 117(2): p. 480-487.
The Expectations
Hours• 10 hours/week in the lab (minimum)• 2-3 hours reading
Log book
• Maintain daily journal• Include objective, all protocols, results, and conclusions
Update • Weekly• Attendance at group meetings; usually on Fridays
Present• Once per semester• 30-40 min
Expand• All project objectives are expandable• Differentiate between B+, A-, and A
Data• Publishable = A+
The Next Steps
• Email your top 3 choices for pre-allocation
• And wait…
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