Process Catalog 2014 - page 30

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Oligonucleotide Purifications
Table V shows the different particle sizes available with the
anion exchange SuperQ functionality, which is typically
used for oligonucleotide purifications. The relative
binding capacities and predicted resolution of the five
particle sizes are depicted by a series of “+” characters.
The more “+” characters listed in the table the better one
resin is relative to another for that parameter. If a process
is developed using one of the five resins and more resolu-
tion is needed, select an appropriate smaller particle size
product. Similarly if more capacity is needed, and resolu-
tion is not a critical issue, a larger particle size resin can be
selected.
The larger particle TOYOPEARL resins are less crosslinked
than the corresponding TSKgel 5PW type resin products
and have more active sites for ligand attachment. Thus they
have higher capacities than the TSKgel 5PW-type resins. In
some cases, TOYOPEARL GigaCap Q-650M (also shown in
Table IV) and its very high capacity can be used, although its
selectivity is somewhat different than the other quaternary.
TSKgel SuperQ-5PW products typically have 2-4 times the
binding capacity of other small particle anion exchange
resins available on the market. This has significant bearing
in the area of difficult to resolve “n-1” DNA and RNAi puri-
fications as loading amounts are increased. Under higher
loading conditions, the TSKgel SuperQ resins maintain
their resolution much better than smaller particle, lower
capacity resins. The smaller particle products may start out
with a slight separation advantage under low oligonucle-
otide loading conditions, but this vanishes as the feedstock
load is increased.
Figure 12 shows a comparison of one smaller particle
size, competitive product, which starts out having better
resolution than TSKgel SuperQ-5PW (20) resin at 1 mg
oligonucleotide/mL of resin. At 20 mg oligonucleotide/mL of
resin, however, the resolution of peaks on the competitive
product deteriorates significantly. The TSKgel SuperQ-5PW
(20) retains excellent resolution even at this higher oligo-
nucleotide level.
Bead size Binding capacity Resolution Bead type Ligand attachment
(mean µm)
TSKgel SuperQ-5PW (20) 20 ++ +++++ methacrylic Type A
TSKgel SuperQ-5PW (30) 30 ++ ++++ methacrylic Type A
TOYOPEARL SuperQ-650S 35 ++++ +++ methacrylic Type A
TOYOPEARL SuperQ-650M 65 ++++ ++ methacrylic Type A
TOYOPEARL SuperQ-650C 100 ++++ + methacrylic Type A
TOYOPEARL GigaCap Q-650M
TOYOPEARL GigaCap Q-650S
75
35
+++++
+++++
++
++++
methacrylic
methacrylic
Type B
Type B
Bead size Binding capacity Resolution Bead type Ligand attachment
(mean µm)
TSKgel SP-5PW (20) 20 ++ +++++ methacrylic Traditional
TSKgel SP-5PW (30) 30 ++ ++++ methacrylic Traditional
TOYOPEARL SP-650S 35 ++++ +++ methacrylic Traditional
TOYOPEARL SP-650M 65 ++++ ++ methacrylic Traditional
TOYOPEARL SP-650C 100 ++++ + methacrylic Traditional
TOYOPEARL GigaCap S-650M
TOYOPEARL GigaCap S-650S
75
35
++
++++
methacrylic
methacrylic
Type B
Type B
+++++
+++++
Oligonucleotide Purification Products:
Peptide Purification Products:
Tosoh Bioscience Products for Peptide Synthesis:
TSKgel SP-3PW (30) 30 +++ ++++ methacrylic Traditional
TABLE V
Ion exchange
chromatography