"The latest report in the 4C suggests however, that they have not found a solution that cuts the mustard" agree 100%
Below conclusion from their study clearly shows that traditional tech which is covalent coupling is not cutting it for Philips ( and imagine a lot of others!!).. info care of jak444 ...
"■ CONCLUSIONS
We have quantified the solution based antigen capturing
activity of antibodies immobilized on magnetic nanoparticles,
for a wide range of surface coverages, with cTnI as the model
antigen. The number of active antibodies that can capture
antigen molecules from solution increases with surface coverage
initially and saturates in the monolayer coverage regime.
We
find that only a small fraction (about 4%) of the total
immobilized antibodies is able to capture antigen from solution,
which decreases even more as the antibody coverage increases.
We attribute this result to the
random orientation and possible
unfolding of the immobilized antibodies at low surface
coverage, and to additional crowding effects at higher
coverages. These mechanisms can impair the availability of
Fab fragments to capture antigen molecules"
From the above its clear that MixnGo fixes these issues..
Further more i want to highlight the preparation of the nanoparticles.. its just horrendous when comparing it with MixnGo imvho!!
Functionalization of Magnetic Nanoparticles with Antibodies.
Magnetic nanoparticles (MNP) were funtional- ized with anti-troponin antibodies at different surface coverages. The immobilization conditions, e.g., pH and EDC to antibody ratio, were the result of an optimization process for high antibody activity. Briefly, the MNPs were ashed twice with 15 mM MES buffer (2-(N-morpholino)ethanesulfonic acid, pH 6.0) by holding the particles with a magnet and resuspended in MES buffer. The nanoparticle surface was then activated by incubation with EDC (N-3-(dimethylamino)propyl-N-ethyl- carbodimide hydrochloride) and sulfo-NHS (N-hydroxysulfo- succinimide) at an end concentration of 2 mg mL−1 (12.8 μM) and 0.5 mg mL−1 (2.3 μM) in an incubator shaker (1000 rpm) at room temperature.
After incubation with EDC and sulfo- NHS for 30 min, the particles were separated by a magnet and
washed twice with MES buffer. The antibody solution was then quickly added to the activated particles. The monoclonal anti- troponin antibodies (MAb1) were added to a final antibody-to- particle mass ratio ranging from 5 to 100 μg of ntibody per 1 mg of nanoparticles. Antibodies and particles were incubated under continuous stirring (1000 rpm)
at room temperature for 30 min. After incubation, the supernatant was removed by holding the particles with a magnet and he particles were
further incubated with Tris−HCl solution (50 mM, pH 7.4)
for 30 min to deactivate the unbound carboxyl groups on the particle surface. This was
followed by two washing steps with storage buffer (Ademtech, France).
Finally, the antibody coupled n
anoparticles were resuspended in storage buffer at an end concentration of 10 mg mL−1
and stored at 4 °C after sonication. Samples were also sonicated briefly after each magnetic separation step performed in the antibody coupling protocol. The loss of particles in pipetting and magnetic separation steps was negligible.
Wood and trees again.. pity come just cannot grasp that changing from old to completely new tech is a slow and painful process but one which has great reward for the patient...
