Environmental Engineering Reference
In-Depth Information
The 2-DE remains the most widely used proteomic technique owing to the fact
that it is currently the method that can separate and measure hundreds of proteins
from a single biological sample simultaneously [ 75 ]. Analysis by 2D-PAGE has
been used for proteomic studies of breast cancer [ 76 ], prostate cancer [ 77 ], hepa-
tocellular carcinoma [ 78 ], renal cell carcinoma [ 79 ], rheumatoid arthritis [ 80 ] and
hepatitis B infection [ 81 ]. Proteomic profiling allows for biomarker discovery based
on the differences in protein expression levels between samples that have been
analysed on a broad scale; that is, normal vs. disease model study, protein expres-
sion knockdown by siRNA or other conditions.
15.3.4 As a Tool for Exploring the Targeting Potential
The targeting potential of the nano/micro systems to the MPS cells that serve as
microbial reservoirs can be studied using 2-DE. The adsorption patterns is qualita-
tively and quantitatively assessed. The relative adsorption by various organs can
help to understand the organ distribution of the delivery systems.
The effect of surface (charge) modified submicron emulsions (cationic and
anionic) on in vitro adsorption of plasma proteins via 2D PAGE has been carried
out [ 82 ]. The presence of poloxamer 188 eliminated the adsorption of the larger
proteins like immunoglobulins, fibrinogen, etc. The smaller proteins such as apo-
lipoproteins and albumin were almost completely adsorbed onto the submicron
emulsions. The adsorption of apolipoprotein, especially apoA-1, was approxi-
mately three times more on stearylamine- and oleylamine-based cationic emulsions
and oleic acid-based anionic emulsions. In addition, the ratio between the apoA-1
and apoA-IV was found to be 1 for lipofundin MCT 10 % whereas it was about 0.26
for deoxycholic acid-based anionic emulsion and above 5 for oleic acid based
anionic emulsions and cationic emulsions. This indicated that emulsions having
similar surface/interfacial charge imparted by different anion-forming stabilisers
(oleic or deoxycholic acids) exhibited markedly different protein adsorption
patterns.
Shegokar et al., identified protein adsorbed on nanocrystals of nevirapine (anti-
retroviral drug) in vitro to predict in vivo fate of nanoparticle [ 83 ]. Protein pattern
analysed showed macrophage targeting might be dominant for polysaccharide
coated nanocrystals and brain targeting for albumin coated nanocrystals
(Fig. 15.6 ). This predicted hypothesis was confirmed by in vivo results obtained.
This type of prediction helps batch selection for in vivo experiments, thereby
reducing the number of animal experiments required, time and cost as well. The
data obtained on nevirapine confirm in vitro and in vivo correlation. Didanosine
NLC showed targeting potential to brain when tested by 2D PAGE technique and
can be used for HIV related dementia conditions and evaluation of the in vitro
differential protein adsorption patterns of didanosine-loaded nanostructured lipid
carriers (NLCs) for potential targeting to the brain [ 84 ].
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