GE HEALTHCARE AKTAprime plus, Краткое руководство по эксплуатации

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11-0027-48 AC, 2007-09 • p35
Ordering information
Product Quantity Code No.
HiTrap SP HP 5 × 1 ml 17-1151-01
HiTrap SP FF 5 × 1 ml 17-5054-01
HiTrap CM FF 5 × 1 ml 17-5056-01
HiTrap SP XL 5 × 1 ml 17-5160-01
HiTrap Capto S 5 x 1 ml 17-5441-22
HiTrap Capto MMC 5 x 1 ml 11-0032-73
HiTrap IEX Selection Kit 7 × 1 ml 17-6002-33
HiTrap Desalting 5 × 5 ml 17-1408-01
100 × 5 ml
*
11-0003-29
HiPrep 26/10 Desalting 1 (
53 ml)
17-5087-01
4
(53 ml)
17-5087-02
Superloop 10 ml 1 18-1113-83
Superloop 50 ml 1 18-1113-84
Superloop 150 ml 1 18-1023-85
* Pack size available by special order
Troubleshooting
High backpressure:
•
Column clogged
– Clean the column according
to instructions. Make sure the sample has been
centrifuged and/or filtered through a 0.45 µm filter.
•
System clogged – Replace the column with a piece
of tubing. Check pressure. If backpressure > 0.3 MPa,
clean system according to manual.
No binding:
•
Check that the correct column is used.
•
Check that the inlet tubing from each buffer is
connected to the correct inlet port.
•
Check that the composition and pH of the buffers are
correct. If the compostion and pH of the buffers are
correct, but there is still no binding:
a) If the protein of interest does not bind to the
column, the pH should be decreased.
b) If it still not binds, it is advisable to try an anion
exchanger (see cue card: Anion exchange).
c) If the protein of interest binds to the column but
the separation is poor, the pH should be increased.
•
Check that the sample has been adjusted to start
buffer conditions.
•
Check that your sample contains target protein.
No elution:
•
Check that the inlet tubing from each buffer is
connected to the correct inlet port.
•
Check that the composition and pH of the buffers are
correct.
•
Use alternative elution conditions according to the
column instructions.
•
Check that your sample contains target protein.
0
AU
280nm
0
20
40
60
80
% B
0.1
0.2
0.3
0.4
0 10 20 30
–– UV 280 nm
––
Programmed %B
Inject
min
15 25
//
Sample: Protein mix containing chymotrypsinogen A, cytochrome C
and lysozyme in start buffer
Column: HiTrap SP HP 1 ml
Start buffer (A1): 20 mM sodium acetate, pH 5.5
Elution buffer (B): 20 mM sodium acetate, 1.0 M NaCl, pH 5.5
5
Typical result