SURFACE IMMOBILIZATION AND

CLUSTERIZATION ADSORPTION PROCESS

OF ANTI C1Q MONOCLONAL ANTIBODY ON
SOLID SURFACES. ATOMIC FORCE
MICROSCOPY INVESTIGATION

CONCLUSION
The results show that the clusteralization process depends on the
surface-protein interactions, as well as protein-protein interactions We
have determined that C1q antibody adopts a flat-on orientation once
adsorbed on the solid surfaces. The ANTB tends to aggregate, forming
clusters as evidenced by atomic force microscopy, particularly at pH 4
when ANTB is positively charged. Hydrophobic glass inhibits the
formation of ANTB clusters at isoelectric pH 5.5. Positively charged
monoclonal antibodies have the highest tendency for clusteralization,
despite electrostatic repulsive interactions, suggesting that other
intermolecular forces, such as hydrogen bonds are prevalent. Using
various ionic strengths it appears that the electrostatic intermolecular
interactions are not important for this aggregation process within
ANTB adsorbed layer. They may play a role in the conformational
changes influencing the exposure of specific aminoacids residues
involved in hydrogen bonds formation. We suggest that the long
distance hydrogen bonds are the most important intermolecular
forces involved in the clusteralization process of the adsorbed
monoclonal antibodies. Clearly, the data strongly suggest that
intermolecular protein-protein forces, as well as protein-surface
interactions have a paramount importance over the preservation of
biological activity of the adsorbed antibody molecules.

SELF ASSEMBLY OF AN ANTISENSE

DIGOXIGENIN LABELED
OLIGONUCLEOTIDE
CORRESPONDING TO NMDA
NEURONAL RECEPTOR

ABSTRACT
The major goal of this work is to study the self assembly of an
antisense digoxigenin labeled oligonucleotide (ADLO)
corresponding to NMDA neuronal receptor. We have used a
molecular probe consisting of ADLO corresponding to NMDA
neuronal receptor, namely to subunit NR1. We have chosen to
study this target, due to the importance of NMDA receptor in
various neurological and psychiatric disorders, like
Parkinson`s disease, Alzheimer`s disease and schizophrenia.
The molecular probe ADLO was dissolved in borate buffer, pH 8
and immobilized on the following surfaces: graphite, silica,
mica and gold. The AFM observations and FTIR results show
that surface immobilization has a paramount importance, due
to the specific interactions through hydrogen bonds among
ADLO molecules and their interaction with the solid substrates.
Anyhow, on either surface the forming of secondary structures
and clusterization is observed. These data are suggesting that
intermolecular hydrogen bonds formation is important for
creating the secondary structure networks. These data open
the possibility for the design of better experimental conditions
for in situ hybridization experiments, in order to avoid
aggregations of ADLO, that affect the affinity for the main
target, respectively mRNAs species.

CONCLUSIONS

In the presence of Mg ions, the binding of DNA to mica is very

strong, the DNA molecules are well adsorbed and could be
easily observed. However, DNA molecules adopt a very
compact structure with many kinks and nodes due to intrastrand base pairing.
On a APTES modified mica surface, the binding of DNA is very
strong. The structure of DNA is compact, with many loops and
intra-strand base pairing. Mica in the presence of Mg ions has a
high affinity for different DNA species. However, the formation
of secondary structures is almost inevitable.
On the graphite surface adsorption, the DNA molecules do not
form any secondary structure.
Future studies are required to establish the relationship among
oligonucleotides primary sequence , GC content and different
secondary structures formation by AFM experiments on mica
surfaces, which may be useful in designing in situ
hybridisation experiments.