211 Bis Avenue Kennedy - BP 1140
03103 Montluçon cedex - France
tel (33) (0) 4 70 03 88 55
Fax. (33) (0) 4 70 03 82 60
|
Sterilization by Autoclaving |
| KAPTIV-M resin was sterilized by autoclaving for up to 30 min. at 120 °C and at neutral pH. Ligand degradation was monitored by RP-HPLC analysis of the sterilizing buffer, but no peaks were observed. KAPTIV-M resin, before and after sterilization procedure, was tested for its ability to purify M Immunoglobulins from crude hybridoma supernatant: data show that the matrix can be autoclaved with no apparent loss of IgM binding and recovery, as determined by ELISA assays. |
|
Experimental conditions |
|
Column volume |
1 ml |
|
Sample |
Crude hybridoma supernatant |
|
IgM loaded |
0.9 mg |
|
Sample volume |
15 ml |
|
Loading buffer |
0.05 M Na-Phosphate, pH 7.0 |
|
Elution buffer |
0.1 M Acetic Acid |
|
Neutralizing buffer |
1.0 M Tris-HCl, pH 9.0 |
|
Regeneration buffers |
M Acetic Acid, 0.5 M NaCl, pH 4.0 0.1 M Tris-HCl, 0.5 M NaCl, pH 8.0 |
|
Results |
|
Resin |
Source |
Antibody |
Total Protein (mg) |
IgM recovery (mg) % |
% Purity SDS-PAGE) |
|
KAPTIV-M |
Crude hybridoma supernatant |
IgM |
45 |
0.9 100 |
> 90 |
|
KAPTIV-M AUTOCLAVED |
Crude hybridoma supernatant |
IgM |
45 |
0.86 96 |
> 90 |
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211 Bis Avenue Kennedy - BP 1140 03103 Montluçon cedex - France tel (33) (0) 4 70 03 88 55 Fax. (33) (0) 4 70 03 82 60 |