Further investigation demonstrated that 3F1 is ideal for affinity purification of rHuIFN α2a, resulting from the recovery rate of 0.8 mg￣1.0 mg of rHuIFN α2a per ml affinity gel with per ml affinity gel the purity of more than 95% and activity of 2×10 11 u/g, and beneficial to the renaturation of rHuIFN α2a.
Results SDS PAGE and Western blot showed that HPV16 L1(m202) protein had been expressed in sf9 cells, the efficiency of purification and renaturation was approximately 17%, the anti serum has the ability of competing inhibition HPV16 McAb reacting with HPV16 L1(m202) protein.
3,The inclusion bodies were dissolved by 6 mol/L guanidine hydrochloride and 50 mmol/L P -mercaptoethanol,then was diluted in the buffer contained 0.5 mol/L Arg to renature for 72 h. The activity experiments verified that the denaturating protein recovered its bioactivity.
Secondly, the monoeric rhTGF-β3 were added to a special solution containing of 1mol/L NaCl, 05mol/L l-Arg, 0.5mmol/L GSSG, 30mmol/L CHAPS and 20%(v/v)DMSO to renature. Finally,the renatured rhTGF-β3 was purified by a DEAE Sepharosel Fast Flow column and a Sephacryl S-100 column.
Results: The expression vector pET28aHsp65E6/E7 was constructed successfully and E6/E7 gene was mutated correctly. Hsp65E6/E7 fusion protein was renatured and purified on the affinity chromatography column simultaneously. The protein purity achieved 95% after the anionic exchange chromatograph purification.
RESULTS Dialysis, anion exchange chromatography and gel filtration renatured ETI with relative renaturation ratios of 20.1%, 32.1% and 50.8% and relative recoveries of protein 16.6%, 15.1% and 55.2%,respectively.
The biological activities of renatured IL13 fusion protein could reach 80～120 U/ml and the fusion proteins could be cut into 26 kD GST and 10 kD IL13 with the treatment of thrombin for several hours.
RESULTS The final concentration of fibrin in separated gel was 0.4×10~(-3) mg·L~(-1). After electrophoresis, zymogram was obtained when the gel was renatured with 2.5% TritonX-100 for 30 min, incubated with PBS buffer(pH 7.2) for 30 min, stained with Comassie Brilliant Blue R-250, restained with 7% acetic acid.
Immunochemical properties of recombinant A-subunit expressed in Escherichia coli and renatured were investigated using a panel of monoclonal antibodies raised against three mistletoe toxins (MLI, MLII, and MLIII).
The recombinant protein PKPI-B10 obtained as inclusion bodies was denatured, separated from admixtures by ion-exchange fast protein liquid chromatography (FPLC) on MonoQ under denaturing conditions, and renatured.
Although the oxidized and reduced glutathione (GSSG and GSH) were employed in the oxidized mobile phase, the reduced-denatured insulin still could not be renatured.
The three disulfide bridges of insulin were formed correctly and the reduced-unfolded insulin can be renatured completely.
The renatured protein purified by Ni-NTA Superflow resins showed ability to bind to antigen on SPC-A-1 lung adenocarcinoma.