LBL-assembly is an established method for the fabrication of multi-composite ultrathin films on solid

surfaces (Decher, et al., 1992). Typically, this technique is based on the use of polyelectrolytes of
opposite charges assembled layer-wise on the surface of interest as shown in Figure 1, thereby building
up a layer system of tunable characteristics, in terms of composition, nanometer range thickness, surface
charge, permeability and elasticity (Tan, et al., 2008).

Figure 1: Schematic representation of LBL deposition of oppositely charged polyelectrolytes on a negatively

charged colloidal substrate. Steps 14, including adsorption of a polycation then a polyanion and
washing after each adsorption step, represent the basic build-up sequence for the development of simplest
film. Repetition of these steps result in a colloid coated with multilayers of polyelectrolytes. Finally, there
is possibility of dissolving the core resulting in capsule formation.

While LBL-assembly is easy to envision on macroscopic objects, it is much more difficult to imagine
how very small objects that cannot be handled individually can be coated. In the case of very small
objects, it is required to bring the objects in question in contact with an oppositely charged
macromolecule in the same solution, a situation that is well known as a classic condition for bridging
flocculation as shown in Figure 2 (Qiu, et al., 2001).

Figure 2: Schematic showing bridging flocculation of two nanoparticles connected by a single chain of long
polymer (Left) and two nanoparticles wrapped individually with short polymers that are in stoichiometric
excess (Middle). An electron micrograph shows a suspension of flocculated nanoparticles (Right).

The power of LBL technology to functionalize nanoparticles smaller than 100 nm while limiting
aggregation and bridging of nanoparticles was first introduced in 1999 by Caruso. Since then, coating of
nanoparticles by LBL technology is an active area of research and is currently considered a hot topic with
many potential applications in drug delivery (Antipov & Sukhorulov, 2004). Due to the versatility of the
LBL surface modifications, Schneider proposed an ideal design for a nanoparticle-based core/shell system
using gold (Schneider, et al., 2009). The core-shell gold nanoparticles (AuNPs) for nucleic acid delivery
consist of four polymer layers deposited on a gold core (Figure 3). Gold was chosen as the size template
for several reasons: The coating process can be monitored because of a red shift of the plasmon
absorption band (Goodman, et al., 2004). It is therefore also possible to identify particle aggregation
during polymer deposition. Further advantages of AuNPs include their limited cytotoxicity and are visible
by TEM with a high contrast because of their large electron density (Connor, et al., 2005).

Figure 3: Schematic representation of nanoparticles coated with multilayer shells as an ideal drug delivery system
(Schneider, et al., 2009).

The first layer is the stabilizing agent mercaptoundecanoic acid (MUA) which conserves a negative
charge on the particle surface (Gittins & Caruso, 2001) as shown in Figure 4, The positively-charged
layer of poly(ethylenimine) (PEI) is followed by the negatively-charged nucleic acid. The nucleic acids
are protected by a last layer of PEI (Elbakry, 2009).

Figure 4: Schematic figure of a gold nanoparticle core stabilized by MUA. The thiol groups displace the citrate
ligands due to their high affinity to the gold surface. The negative charge of the deprotonated carboxylic
acid group conserves a negative charge on the particle surface which is the basis for the further
deposition of positively-charged polymers and prevents aggregation by electrostatic repulsion.

Figure 5: Schematic illustration of the LBL process. The stabilized gold nanoparticles are alternatively covered
with PEI and DNA to give the LBL-assembled core-shell particles for nucleic acid delivery into
mammalian cells (Elbakry, 2009).

Hence, with the functionalization of AuNP using LBL technology as shown in Figure 5, a novel gene
carrier system to transport nucleic acids (DNA or RNA) into mammalian cells for the treatment of genetic
diseases can be fully developed without a need to worry about the degradation of nucleic acid.