Journal: Microbial Cell Factories

Article Title: Secretion of tumoricidal human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) by recombinant Lactococcus lactis : optimization of in vitro synthesis conditions

doi: 10.1186/s12934-018-1028-2

Figure Lengend Snippet: Lactococcus lactis -derived hsTRAIL-induced death of human colon cancer HCT116 cells in vitro. HCT116 cells were incubated for 48 h with increasing concentrations of hsTRAIL in supernatants from broth culture of L. lactis (hsTRAIL+). The following controls in experimental setup were used: corresponding volumes of supernatants of L. lactis (hsTRAIL−)—negative control; corresponding concentrations of recombinant human TRAIL (Peprotech)—positive control. Viability of cancer cells was assessed by MTS assay. Results are presented as % of viability of cells incubated in standard culture medium only. hsTRAIL present in supernatant from L. lactis (hsTRAIL+) broth culture decreased viability of cancer cells in a dose-dependent manner, comparable to corresponding concentrations of positive control. Table below—concentration of TRAIL [ng/ml] in specified volume of supernatant or concentration of standard TRAIL formulation (rhTRAIL, Peprotech). The bars indicate the mean value ± SD of three independent experiments, each performed in triplicates. Statistical significance was calculated using two-way ANOVA with Tukey’s multiple comparison post-test. *p < 0.05, **p < 0.01, ***p < 0.001

Article Snippet: As positive control, standard TRAIL formulation (rhTRAIL, Peprotech) in corresponding concentration was used ( e ).

Techniques: Derivative Assay, In Vitro, Incubation, Negative Control, Recombinant, Positive Control, MTS Assay, Concentration Assay, Formulation, Comparison

Journal: Microbial Cell Factories

Article Title: Secretion of tumoricidal human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) by recombinant Lactococcus lactis : optimization of in vitro synthesis conditions

doi: 10.1186/s12934-018-1028-2

Figure Lengend Snippet: Induction of apoptosis of colon cancer HCT116 cells by hsTRAIL from supernatant of L. lactis (hsTRAIL+). HCT116 cells were treated with supernatants from broth cultures of L. lactis (hsTRAIL±) bacteria and rhTRAIL (positive control) in a corresponding dose of 100 ng/ml for 48 h and then stained with FITC-Annexin V and PI to determine cancer cells apoptosis. HCT116 turned apoptotic after treatment with supernatant of L. lactis (hsTRAIL+) bacteria ( a , d ) in contrast to supernatant of L. lactis (hsTRAIL−) (negative control; c ) or medium ( b ). As positive control, standard TRAIL formulation (rhTRAIL, Peprotech) in corresponding concentration was used ( e ). Dot-plots. FL1-H—FITC; FL2-H—PI. The bars ( a ) indicate the mean value ± SD of three independent experiments, each performed in triplicates. Differences between the groups were calculated using two-way ANOVA with Tukey’s multiple comparisons post-test. *p < 0.05

Article Snippet: As positive control, standard TRAIL formulation (rhTRAIL, Peprotech) in corresponding concentration was used ( e ).

Techniques: Bacteria, Positive Control, Staining, Negative Control, Formulation, Concentration Assay

Journal: Biotechnology progress

Article Title: A Simplified Method for the Efficient Purification and Refolding of Recombinant Human TRAIL

doi: 10.1002/btpr.3007

Figure Lengend Snippet: Expression, purification and refolding of recombinant human TRAIL (rhTRAIL).

Article Snippet: Cells were incubated with purified rhTRAIL and commercial rhTRAIL (Enzo Life Science) at 5, 10, 20 and 40 ng/mL for 2 hours before being collected and stained with Annexin V and Propidium Iodide (PI).

Techniques: Expressing, Purification, Recombinant

Journal: Biotechnology progress

Article Title: A Simplified Method for the Efficient Purification and Refolding of Recombinant Human TRAIL

doi: 10.1002/btpr.3007

Figure Lengend Snippet: Samples were analyzed in 12% Tris-Tricine SDS-PAGE and stained with Coomassie Bright Blue. Lane 1: molecular weight marker. Lane 2: total cell lysate. Lanes 3: fraction washed off the Ni-NTA column by the lysis buffer containing 30 mM imidazole. Lane 4: fraction of purified rhTRAIL eluted by the elution buffer containing 300 mM imidazole. The arrow indicates the rhTRAIL protein.

Article Snippet: Cells were incubated with purified rhTRAIL and commercial rhTRAIL (Enzo Life Science) at 5, 10, 20 and 40 ng/mL for 2 hours before being collected and stained with Annexin V and Propidium Iodide (PI).

Techniques: SDS Page, Staining, Molecular Weight, Marker, Lysis, Purification

Journal: Biotechnology progress

Article Title: A Simplified Method for the Efficient Purification and Refolding of Recombinant Human TRAIL

doi: 10.1002/btpr.3007

Figure Lengend Snippet: All the samples were diluted 10 times and analyzed in 12% Tris-Tricine SDS-PAGE and stained with Coomassie Bright Blue. Lane 1: molecular weight marker. Lane 2: denatured rhTRAIL purified by IMAC. Lanes 3: rhTRAIL renatured by stepwise dilution. Lane 4: rhTRAIL after dialysis to remove urea completely. Lane 5: rhTRAIL after concentration with protein concentrator. The arrow indicates the TRAIL protein.

Article Snippet: Cells were incubated with purified rhTRAIL and commercial rhTRAIL (Enzo Life Science) at 5, 10, 20 and 40 ng/mL for 2 hours before being collected and stained with Annexin V and Propidium Iodide (PI).

Techniques: SDS Page, Staining, Molecular Weight, Marker, Purification, Concentration Assay