Table 1 |
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|
Recovery of Tat protein and removal of endotoxin |
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|
Tat Sample |
Endotoxin (EU/μg) |
Yield (mg/Liter) |
||
|
|
||||
|
B-Tat |
C-Tat |
B-Tat |
C-Tat |
|
|
|
||||
|
Neat E. coli lysate |
340 |
320 |
- |
- |
|
Ni-NTA (without Triton wash) |
7.6 |
5.5 |
3.0 |
5.0 |
|
Ni-NTA (after Triton wash) |
0.75 |
0.55 |
2.0 |
4.0 |
|
After SP-Sepharose |
0.039 |
0.034 |
0.5 |
1.0 |
|
|
||||
|
Tat protein recombinantly expressed in bacteria was purified using Ni-NTA column chromatography and ion-exchange column chromatography performed sequentially. Protein concentration of the eluted Tat protein was determined using a dye binding assay [99]. Endotoxin concentration in the samples was determined using a commercial kit (QCL-1000, Biowhittaker). |
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|
Siddappa et al. Retrovirology 2006 3:53 doi:10.1186/1742-4690-3-53 |
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