Table 1. Protein-ligand (protein A) immobilization protocols performed in accordance with the manufacturer’s recommendations for the CM5 chip and for the CMD500M chip.
|
# |
Procedure |
CM5 |
CMD500M |
|
1 |
Surface activation |
7 min, 10 ml/min, 0.2 М EDC/0.05 М NHS |
10 min, 5 ml/min, 0.2 М EDC/0.05 М NHS |
|
2 |
Protein ligand immobilization |
7 min, 5 ml/min, ligand 100 mg/ml in 10 mМ acetate buffer (рН 4.5) |
10 min, 5 ml/min, ligand 100 mg/ml in 10 mМ acetate buffer (рН 4.5) |
|
3 |
Surface inactivation |
7 min, 10 ml/min, 1 М ethanolamine -HCl (pH 8.5) |
15 min, 5 ml/min, 1 М ethanolamine -HCl (pH 8.5) |
|
4 |
Running buffer |
HBS-EP+ (рН 7.4) |
dd H2O |
|
5 |
Level of protein ligand immobilization |
3000 RU |
3400 RU |