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FranciGen specializes on the
the development and sales of application specific capturing schemes for
proteins, antibodies, DNA and other natural substances present in dilute
concentrations in untreated solutions such as lysates.
The capturing process is
based on affinity precipitation with thermo-responsive polymers. |
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Affinity
Precipitation
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Affinity precipitation
is based on the platform technology "stimulus-responsive polymers", also
called "smart materials". These polymers show a highly nonlinear behavior
towards external stimuli, such as the temperature. When a certain temperature
is surpassed, polymers, which were perfectly water-soluble before, will
flock out (precipitate) within seconds. This process is fully
and quickly reversible, and such polymers can be precipitated, re-dissolved
and precipitated again many times by minor changes of the temperature.
In order to exploit this effect for product isolation, it suffices to
chemically conjugate the polymer to a small affinity ligand, which is
capable of specifically recognizing the target molecule. The result is
the so-called affinity macroligand (AML), a molecule, which can be used
to first bind and subsequently co-precipitate the product and only the
product even from most complex raw solutions.
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Thermo-responsive AML
Thermo-responsive AML are water-soluble poly-N-alkylacrylamides with defined
chain length, transformation temperature and terminal affinity end group.
The affinity end group can be varied according to need and allows the
easy capture of a variety of target molecules. The AML have oligomeric
character (short chain length), hence assuring (i) very homogeneous structure
and size, (ii) uniform affinity environment, (iii) high ligand efficiency,
(iv) optimum ligand accessibility, and (v) lack of non-specific interaction.
Only these five features guarantee the specific and efficient purification
of the target substance. This thermoresponsive effect is independent
of the conjugated ligand and therefore a wide variety of such ligands
can be applied.
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The liquid
(for example cell lysate made from bacteria) containing the target molecule
is mixed with an appropriate amount of affinity macroligands (AML) under
appropriate affinity conditions. The AML is a conjugate between our dedicated
oligomeric AML-precursor and a ligand, whose affinity is specific to a
motif on the target molecule.
By increasing
the temperature the AML-Target complex is precipitated, typically in the
presence of a suitable filter aid.
After filtration
and washing, the specifically bound target molecule is released by washing
the complex with a dissociation buffer. Filter aid and uncharged AML can
be reused. |