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…