Journal: Vaccines

Article Title: Challenges and Opportunities in the Process Development of Chimeric Vaccines

doi: 10.3390/vaccines11121828

Figure Lengend Snippet: List of commercially approved human and veterinary vaccines developed using an insect cell expression system.

Article Snippet: Classical Swine Flu Fever Virus , Bayovac ® CSF E2 , E2 protein/Subunit , Sf-21 cells , BAYER AG/Pfizer Animal Health (Nordrhein-Westfalen, Germany/Groton, CT, USA (Discontinued) , [ ] .

Techniques: Vaccines, Expressing, Virus

Journal:

Article Title: The Nramp orthologue of Cryptococcus neoformans is a pH-dependent transporter of manganese, iron, cobalt and nickel

doi: 10.1042/BJ20040836

Figure Lengend Snippet: (A, B) Sf 21 cells infected with recombinant baculovirus encoding Cramp–Myc, immunostained with an anti-Myc antibody (FITC channel and light-field images respectively). (C, D) Control cells infected with wild-type baculovirus. Not all cells are expressing Cramp, as only approx. 70% of cells in the monolayer were infected by recombinant baculovirus.

Article Snippet: Sf 21 cell transfection and recombinant virus clone isolation Spodoptera frugiperda Sf 21 cells (BD Biosciences, Clontech) were co-transfected with linearized baculovirus DNA (BD Biosciences, Clontech) together with pBacPAK8-Cc#2, according to the BacPAK™ Baculovirus expression system user manual (BD Biosciences, Clontech) [ 25 ].

Techniques: Infection, Recombinant, Expressing

Journal:

Article Title: The Nramp orthologue of Cryptococcus neoformans is a pH-dependent transporter of manganese, iron, cobalt and nickel

doi: 10.1042/BJ20040836

Figure Lengend Snippet: (A) Mn2+ accumulation at pH 5.5 by Cramp-transfected Sf 21 cells compared with control cells (WT) infected with non-recombinant baculovirus (total [Mn2+] 1 μM). Mn2+ uptake is expressed in units of pmol/h per 106 cells. Cramp induced up to 38-fold increases in Mn2+ uptake compared with controls. Cramp-mediated Mn2+ uptake is approximately linear over the time frame 0–15 min. (B) Mn2+ accumulation by Cramp-expressing Sf 21 cells (pH 5.5) as a function of total Mn2+ concentration. The inset shows a Lineweaver–Burk plot of these data. A Km of 24±4 μM and a Vmax of 1700±102 pmol/h were estimated by non-linear regression. (C) pH profiles for Mn2+ transport by Cramp-expressing and control (WT) Sf 21 cells. Essentially all Mn2+ transport between pH 5.0 and 6.0 is mediated by Cramp. At pH 6.5 and above, endogenous Mn2+ transport is apparent in control cells. Superimposition of the curves for Cramp-transfected and control cells shows that Cramp does not contribute any significant additional Mn2+ uptake at pH 6.5 and above, indicating that Cramp-mediated Mn2+ uptake is abrogated at pH values in excess of 6.5. Maximal Cramp activity was seen at pH 5.5–6.0. Mn2+ transport at pH 5.0 was significantly lower than at pH 5.5/6.0 (P=0.05). Error bars show S.E.M.

Article Snippet: Sf 21 cell transfection and recombinant virus clone isolation Spodoptera frugiperda Sf 21 cells (BD Biosciences, Clontech) were co-transfected with linearized baculovirus DNA (BD Biosciences, Clontech) together with pBacPAK8-Cc#2, according to the BacPAK™ Baculovirus expression system user manual (BD Biosciences, Clontech) [ 25 ].

Techniques: Transfection, Infection, Recombinant, Expressing, Concentration Assay, Activity Assay

Journal:

Article Title: The Nramp orthologue of Cryptococcus neoformans is a pH-dependent transporter of manganese, iron, cobalt and nickel

doi: 10.1042/BJ20040836

Figure Lengend Snippet: (A) Effects of potential bivalent cation competitors on Cramp-mediated Mn2+ transport in Sf 21 cells. Cramp-transfected cells were incubated for 15 min at pH 5.5 in the presence of 1 μM total Mn2+ and a 100-fold excess (100 μM) of the indicated non-radioactive bivalent cation chloride salts. Results are expressed as a percentage of the uptake seen with 1 μM Mn2+ in the absence of competitor. There were no significant differences in uptake between competitor conditions within each bracketed group. (B) Inhibition curves for Fe2+ and Cd2+. Mn2+ transport at pH 5.5 (1 μM total Mn2+) was measured at a series of Fe2+ and Cd2+ concentrations and expressed as a percentage of the Mn2+ transport observed in the absence of the competitor. Negligible endogenous Mn2+ transport occurs at pH 5.5 in Sf 21 cells. (C, D) Cramp-mediated Co2+ and Ni2+ transport. Transport was measured at pH 5.5 in the presence of 1 μM total Co2+ or Ni2+. WT, control cells infected with non-recombinant baculovirus.

Article Snippet: Sf 21 cell transfection and recombinant virus clone isolation Spodoptera frugiperda Sf 21 cells (BD Biosciences, Clontech) were co-transfected with linearized baculovirus DNA (BD Biosciences, Clontech) together with pBacPAK8-Cc#2, according to the BacPAK™ Baculovirus expression system user manual (BD Biosciences, Clontech) [ 25 ].

Techniques: Transfection, Incubation, Inhibition, Infection, Recombinant