Montage Antibody Purification Kits
Montage Antibody Purification Kits are designed for fast
and easy antibody purification from serum, ascites or cell culture
supernatants. The kits include spin columns pre-packed with high capacity
PROSEP-A or G media. There are no pumps to setup. No columns to pack. All you
need is a centrifuge to purify 10–20 mg in less than 60 minutes.
The
protocol is easy to follow and, unlike traditional gravity flow methods, the
use of centrifugal purification allows for walk-away convenience.
Results show that the spin columns can be regenerated up to 10X with no loss
of binding capacity or reduced
specificity.
Performance
Spin Column Reuse
)
Human IgG was purified 10 times from normal serum using a regenerated PROSEP-G spin column.
Purification of MAbs
)
Monoclonal mouse IgG was purified with a PROSEP-A spin column from ascites. (Lane 1: unpurified ascites. Lane 2: flow-through ascites. Lane 3: wash flow-through. Lane 4: 5 µg purified antibody.) Courtesy Exalpha Biologicals.
| Binding Affinities of Protein A and Protein G | ||||||
| Protein | A | G | Protein | A | G | |
| Human IgG1 | ++ | ++ | Rat IgM | + /-- | - | |
| Human IgG2 | ++ | ++ | Rabbit IgG | ++ | ++ | |
| Human IgG3 | -- | ++ | Hamster IgG | + | ++ | |
| Human IgG4 | ++ | ++ | Guinea Pig IgG | ++ | + | |
| Human IgA | + | -- | Bovine IgG | + | + | |
| Human IgD | + | -- | Sheep IgG | + / -- | + | |
| Human IgE | + | -- | Goat IgG | + /-- | + | |
| Human IgM | + | -- | Pig IgG | ++ | ++ | |
| Mouse IgG1 | + | + | Chicken IgG | -- | + /-- | |
| Mouse IgG2a | ++ | ++ | Fragments | |||
| Mouse IgG2b | ++ | ++ | Human Fab | + | + | |
| Mouse IgG3 | + | ++ | Human F(ab')2 | + | + /-- | |
| Mouse IgM | + /-- | -- | Human ScFv | + | -- | |
| Rat IgG | ++ | ++ | Human Fc | + | -- | |
| Rat IgG1 | + /-- | + | Human K | -- | -- | |
| Rat IgG2a | + /-- | ++ | Human | -- | + | |
| Rat IgG2b | + /-- | + | ||||
| Rat IgG2c | + /-- | + | ||||
| Legend: ++ = Strong affinity; + = Moderate/slight affinity; +/-- = Requires evaluation; -- = No affinity | ||||||
The kit includes Montage Spin Columns pre-packed with PROSEP-A or PROSEP-G resin, optimized buffers, Steriflip filter devices for prefiltration and Amicon Ultra centrifugal devices for desalting and concentration of purified antibodies. Montage Spin Columns are also available separately.
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