Refolding, purification, and characterization of constitutive-active human-Smad8 produced as inclusion bodies in ClearColi® BL21 (DE3)

2021 ◽  
Vol 184 ◽  
pp. 105878
Author(s):  
Carla Lizbeth Segovia-Trinidad ◽  
Bastian Quaas ◽  
Zhaopeng Li ◽  
Antonina Lavrentieva ◽  
Yvonne Roger ◽  
...  
Keyword(s):  
Inclusion Bodies ◽  
Purification And Characterization

Expression, purification and characterization of a full-length recombinant HIV-1 Vpu from inclusion bodies

2016 ◽  
Vol 128 ◽  
pp. 109-114 ◽  
Author(s):  
Zikhona Njengele ◽  
Ronel Kleynhans ◽  
Yasien Sayed ◽  
Salerwe Mosebi
Keyword(s):  
Inclusion Bodies ◽  
Full Length ◽  
Purification And Characterization ◽  
Hiv 1

Refolding, Purification, and Characterization of a Loop Deletion Mutant of Human Bcl-2 from Bacterial Inclusion Bodies

1999 ◽  
Vol 15 (2) ◽  
pp. 162-170 ◽  
Author(s):  
Malcolm Anderson ◽  
David Blowers ◽  
Neil Hewitt ◽  
Philip Hedge ◽  
Alexander Breeze ◽  
...  
Keyword(s):  
Inclusion Bodies ◽  
Deletion Mutant ◽  
Purification And Characterization

Purification and Characterization of Recombinant sTRAIL Expressed in Escherichia coli

2004 ◽  
Vol 36 (2) ◽  
pp. 118-122 ◽  
Author(s):  
Xiao-Xia Xia ◽  
Ya-Ling Shen ◽  
Dong-Zhi Wei
Keyword(s):  
Inclusion Bodies ◽  
Tumor Cell Line ◽  
Single Step ◽  
Dilution Method ◽  
High Concentration ◽  
Purification And Characterization ◽  
E Coli ◽  
Metal Affinity ◽  
Soluble Trail

Abstract As a potential anti-tumor protein, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has drawn considerable attention. This report presented the purification and characterization of soluble TRAIL, expressed as inclusion bodies in E. coli. sTRAIL inclusion bodies were solubilized and refolded at a high concentration up to 0.9 g/L by a simple dilution method. Refolded protein was purified to electrophoretic homogeneity by a single-step immobilized metal affinity chromatography. The purified sTRAIL had a strong cytotoxic activity against human pancreatic tumor cell line 1990, with ED50 about 1.5 mg/L. Circular dichroism and fluorescence spectrum analysis showed that the refolded sTRAIL had a structure similar to that of native protein with β-sheet secondary structure. This efficient procedure of sTRAIL renaturation may be useful for the mass production of this therapeutically important protein.

Purification and characterization of naturally occurring HIV-1 (South African subtype C) protease mutants from inclusion bodies

2016 ◽  
Vol 122 ◽  
pp. 90-96 ◽  
Author(s):  
Sibusiso B. Maseko ◽  
Satheesh Natarajan ◽  
Vikas Sharma ◽  
Neelakshi Bhattacharyya ◽  
Thavendran Govender ◽  
...  
Keyword(s):  
South African ◽  
Inclusion Bodies ◽  
Subtype C ◽  
Purification And Characterization ◽  
Naturally Occurring ◽  
Hiv 1

Expression, Purification and Characterization of Amantadine Receptor in Escherichia coli

2012 ◽  
Vol 161 ◽  
pp. 88-93 ◽  
Author(s):  
Hai Xin Sun ◽  
Li Min Cao ◽  
Hong Lin ◽  
Fang Lv
Keyword(s):  
Inclusion Bodies ◽  
Equilibrium Dialysis ◽  
Molecular Size ◽  
M2 Protein ◽  
Purification And Characterization ◽  
E Coli ◽  
His Tag ◽  
Sds Page ◽  
Fast Flow

In order to obtain large quantities of broadly selective receptor as one diagnose agent to detect amantadine residue, the M2 protein gene with a His-tag was ligated into pET11a and transferred into E. coli BL21 (DE3) cell. The recombinant E. coli was cultured in liquid LB culture. SDS-PAGE result showed the recombinant M2 protein (rM2) was expressed as insoluble inclusion bodies with about 18KDa in molecular size. rM2 protein was further recognized by Western blot and purified by Ni Sepharose 6 Fast Flow and then refolded. The equilibrium dialysis result showed the rM2 protein had the binding constant of 1.1×105, and stoichiometry of 4.2. The above result showed the rM2 has the potential as biological diagnose agent to the detection of amantadine residue.

Purification and characterization of phosphoglucomutase from peas

1994 ◽  
Vol 92 (3) ◽  
pp. 479-486 ◽  
Author(s):  
Cynthia M. Galloway ◽  
W. Mack Dugger
Keyword(s):  
Purification And Characterization

Purification and Characterization of Tissue-Type Plasminogen Activator in Maxillary Mucosa with Chronic Inflammation

1985 ◽  
Vol 54 (02) ◽  
pp. 485-489 ◽  
Author(s):  
Yukiyoshi Hamaguchi ◽  
Masuichi Ohi ◽  
Yasuo Sakakura ◽  
Yasuro Miyoshi
Keyword(s):  
Plasminogen Activator ◽  
Chronic Inflammation ◽  
Gel Filtration ◽  
Potassium Acetate ◽  
Tissue Type ◽  
Purification And Characterization ◽  
Tissue Type Plasminogen Activator ◽  
Urokinase Inhibitor ◽  
Type Plasminogen Activator

SummaryTissue-type plasminogen activator (TPA) was purified from maxillary mucosa with chronic inflammation and compared with urokinase. Purification procedure consisted of the extraction from delipidated mucosa with 0.3M potassium acetate buffer (pH 4.2), 66% saturation of ammonium sulfate, zinc chelate-Sepharose, concanavalin A-Sepharose and Sephadex G-100 gel filtration chromatographies.The molecular weight of the TPA was approximately 58,000 ± 3,000. Its activity was enhanced in the presence of fibrin and was quenched by placental urokinase inhibitor, but not quenched by anti-urokinase antibody. The TPA made no precipitin line against anti-urokinase antibody, while urokinase did.All these findings indicate that the TPA in maxillary mucosa with chronic inflammation is immunologically dissimilar to urokinase and in its affinity for fibrin.

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