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Preparation
of Membrane Affinity Cartridge |
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- Patented Cartridge
Shell
In this work, a new kind of cartridge shell was designed and prepared.
This cartridge contains an adjustable cover from which membrane
bundlings with different thickness can be adopted in the same
cartridge. So that its capacity for the membrane media can be
adjustable to meet different demands.
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The construction scheme of the membrane cartridge for membrane affinity
chromatography
1. Holder. 2. Cover. 3. Gasket. 4. Distribution plate. 5. Affinity
membrane.
6. Bonding/Wraping part.
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- Sealing Methods
(Bonding)
Sealing of the system is one of the most important factors
in the preparation of membrance cartridge, it becomes more serious
when the bundling of many single membranes is adopted. In this
work, a special bonding method was performed. The circumference
with the width of 1 mm on the edge of each individual membranes
was saturated with a elastic glue.  After
removing the excess glue, up to 80 sheets of membrane can be packed
and bonded together with mild pressure. The structure of the cartridge
prepared using this method was appropriate both in sealing and
separation efficiency.
This method is advantagous that the bonded membranes
could be processed easily.
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- Sealing Methods
(Wraping)
The bonding method can be not performed until the membrane
was thoroughly dried and therefore not suitable in case macrobiomolecular
was adopted as affinity ligand. In further work, a new sealing
method is developed in which the membranes are wrapped together
with a elastic sealing tape. Thus, the prepared membranes are
clipped together by a clamp, then wrapped with a sealing film
around the edge of the membrane bundling, the film should cover
a 2 mm width of the circumference of the first and last membranes,
and finally the wrapped bundling of membrane was packed into the
cartridge. This method can be performed immediately after the
reaction and therefore much favourable for the use of biomolecular
as affinity ligand.
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Affinity Cartridges
Made by Bonding (left) and Wraping (right) Methods |
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Uniformity
of Ligands Distribution
In membrane affinity chromatography, wide but short
cartridges (bundle of membranes) are used for separation instead
of the thin and long columns employed in the conventional chromatography.
Consequently, the uniformity of the radial and axial distributions
of the immobilized ligand is expected to affect the efficiency
of the cartridge. The radial and axial ligand profiles were
determined by immobilizing color ligands onto the membrane because
the color distribution can provide some qualitative information
about the uniformity of the ligand distribution in the cartridge.
Reactive Red 2, which is a triazine dye, and BSA were used to
evaluate the uniformity of the immobilized ligands. BSA did
not provide any color by itself, but when immobilized via diazotization,
the immobilized BSA exhibited a vivid color. In contrast to
the process of ligand immobilization, in the uniformity determination
a lower ligand content reacting solution and a shorter reaction
time were required. (For too high ligand contents or long reaction
times, no color differences would have been detectable.) The
following figure indicates a high uniformity in the distribution
for both ligands.
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Ligand Distribution
across the Cellulose Membrane
A1, A2 and A3 represent the Reactive Red 2 distribution on the 1st,
10th and 20th membranes, respectively, in the cartridge.
B1, B2 and B3 represent the BSA distribution on the 1st, 10th and
20th membranes, respectively, in the cartridge.
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