Lab Meeting August 8, 2013

By: Charis Fernandez

Techniques Learned

• Gel Electrophoresis• Cell culture• Nanoparticle preparation• Lyophilization• Cytotoxicity

1 2 3 4 5 6 7

1: Empty rHDL2: Myristoyl valrubicin3: Apo A1 valrubicin4: Thyroglobulin5: Plasma serum6: Bovine serum albumin7: Human HDL

Native PAGE Bis-Tris Gel with Tricine Buffer

Thyroglobulin MW: 660,000 DaBSA MW: 66,500 DaHuman HDL MW: 70,000-360,000 DaEstimated rHDL MW: ~170,000 Da

24 hour run

1 2 3 4 5 6 7

1: Empty rHDL2: Myristoyl valrubicin3: Apo A1 valrubicin4: Thyroglobulin5: Plasma Serum6: Bovine Serum Albumin7: Human HDL

Native PAGE Bis-Tris Gel with Tricine Buffer8 hour run

Thyroglobulin MW: 660,000 DaBSA MW: 66,500 DaHuman HDL MW: 70,000-360,000 DaEstimated rHDL MW: ~170,000 Da

Myr-5A-Cisx Nanoparticle Size

0.1 1 5 10 500

20

40

60

80

100

120

140

160

Myr-cisxEmpty rHDL

Concentration (μM)

% C

ell S

urvi

val

MTT Cytotoxicity

Myr-5A-Cisx and Empty rHDL on SJ-N-KP cell line

Size and NMR to determine percent drug incorporated

Empty Nanoparticle Samples

• 0.5 mg Myr-5A peptide• 1.0 mg Myr-5A peptide• 2.0 mg Myr-5A peptide

Encapsulated 50 µl Cisplatin Derivative Samples

• 0.5 mg Myr-5A peptide• 1.0 mg Myr-5A peptide• 2.0 mg Myr-5A peptide

Future Studies

• Western blot of Neuroblastoma cell lines• Cytotoxicity of Cisplatin derivative on different

Neuroblastoma cell lines• Effect of encapsulated Cisplatin derivative on normal

healthy cells