Ultra-small water-dispersible fluorescent chitosan nanoparticles:
synthesis, characterization and specific targeting
Padmavathy Tallury
a
, Sounitra Kar
a
and Swadeshmukul Santra
a,b,c
*
Suwussa Bamrungsap,
d
Yu-Fen Huang,
d
and Weihong Tan
d
ES1
Materials
Chitosan polymer (low molecular wight), Triton X-100, N-hydroxysuccinimide (NHS),
1-ethyl-3-(3-dimethylaminopropyl carbodiimide hydrochloride) (EDC) were purchased from
Sigma-Aldrich Chemical Co., USA; Fluorescein isothiocyanate (FITC), anhydrous ethanol were
purchased from Fisher Scientific. Dialysis cellulose membrane (MWCO, 6-8 kD) was purchased
from Spectrum Laboratories (Rancho Dominguez, CA). Deoxyribonucleotides and 5’-carboxyl
modifiers were purchased from Glen Research (Sterling, VA). All solvents and reagents were
obtained from Fisher Scientific and were used without further purification. CCRF-CEM cells
(CCL-119 T-cell, human acute lymphoblastic leukemia) and Ramos cells (CRL-1596, B-cell,
human Burkitt’s lymphoma) were obtained from American Type Culture Association, USA. All
of the cells were grown in RPMI-1640 containing 10% fetal bovine serum (FBS) and 100 IU/mL
penicillin-Streptomycin at 37 °C in a humid atmosphere with 5% CO
2
. G25 Sephadex size-
exclusion column (NAP
TM
-5) was procured from Amersham Pharmacia Biotech, USA.
Instrumentation
Degree of deacetylation of chitosan was determined by elemental analysis at the Atlantic
Microlabs, Norcross, GA. JEOL JEM 1011 100kV transmission electron microscope (TEM) was
used to characterize particle size. TEM sample was prepared by placing a drop of the chitosan
nanoparticles on a carbon coated copper grid (400 mesh size) followed by air drying. Particle
size distribution and zeta potential in deionized water was measured by using the Malvern Zeta
Sizer (model: NanoZS) Dynamic Light Scattering (DLS) instrument. The concentration of
activated aptamer and the amount of FITC in FITC-Chitosan conjugate polymer was determined
Supplementary Material (ESI) for Chemical Communications
This journal is © The Royal Society of Chemistry 2009