Expressing:Article Title: Vaccination with Killed but Metabolically Active E. coli Over-expressing Hemagglutinin Elicits Neutralizing Antibodies to H1N1 Swine Origin Influenza A Virus
Article Snippet: Subsequently, the inclusion bodies were refolded as recommended by the manufacture (Amersham Biosciences, Piscataway, NJ). .. To further confirm the protein expression, the lysates of E. coli , BL21(DE3) harboring SD/H1N1-S-OIV HA, GFP, or P. acnes CAMP factor were subjected to 10 % SDS-PAGE, immunoblotted on polyvinylidene fluoride (PVDF) membranes and incubated with sera from mice immunized with KBMA E. coli HA SD/H1N1-S-OIV or a commercial antibody to swine origin influenza A (H1N1) HA (ProSci Incorporated, Poway, CA, USA). fig ft0 fig mode=article f1 Figure 1 caption a4 caption a8 Estimation of the expression level of HA of H1N1-S-OIV in E. coli, BL21 (DE3) (1 × 10 8 CFU) Virus growth A clinical isolate of SD/H1N1-S-OIV and two seasonal influenza A/Brisbane/59/2007 (H1N1) virus (ATCCH1N1) and an A/Brisbane/10/2007 (H3N2)-like virus (ATCCH3N2) were grown in virus growth medium containing Dulbecco's modified Eagle's medium (DMEM) with glutamine, 0.2% bovine serum albumin (BSA), 25 mM Hepes and 2 μg/ml TPCK-trypsin (Sigma Chemical Co., St. Louis, Mo.) and propagated in Madin-Darby canine kidney (MDCK) cells as described in our previous publication ( 31 ). .. Intranasal immunization with KBMA E. coli HA SD/H1N1-S-OIV To create KBMA E. coli, the E. coli BL21(DE3) over-expressing HA of SD/H1N1-S-OIV or GFP was inactivated by UV irradiation at total energy of 7000 J/m 2 by a Spectrolinker (Spectronics, Westbury, NY) ( 23 , 25 , 32 ).
Incubation:Article Title: Vaccination with Killed but Metabolically Active E. coli Over-expressing Hemagglutinin Elicits Neutralizing Antibodies to H1N1 Swine Origin Influenza A Virus
Article Snippet: Subsequently, the inclusion bodies were refolded as recommended by the manufacture (Amersham Biosciences, Piscataway, NJ). .. To further confirm the protein expression, the lysates of E. coli , BL21(DE3) harboring SD/H1N1-S-OIV HA, GFP, or P. acnes CAMP factor were subjected to 10 % SDS-PAGE, immunoblotted on polyvinylidene fluoride (PVDF) membranes and incubated with sera from mice immunized with KBMA E. coli HA SD/H1N1-S-OIV or a commercial antibody to swine origin influenza A (H1N1) HA (ProSci Incorporated, Poway, CA, USA). fig ft0 fig mode=article f1 Figure 1 caption a4 caption a8 Estimation of the expression level of HA of H1N1-S-OIV in E. coli, BL21 (DE3) (1 × 10 8 CFU) Virus growth A clinical isolate of SD/H1N1-S-OIV and two seasonal influenza A/Brisbane/59/2007 (H1N1) virus (ATCCH1N1) and an A/Brisbane/10/2007 (H3N2)-like virus (ATCCH3N2) were grown in virus growth medium containing Dulbecco's modified Eagle's medium (DMEM) with glutamine, 0.2% bovine serum albumin (BSA), 25 mM Hepes and 2 μg/ml TPCK-trypsin (Sigma Chemical Co., St. Louis, Mo.) and propagated in Madin-Darby canine kidney (MDCK) cells as described in our previous publication ( 31 ). .. Intranasal immunization with KBMA E. coli HA SD/H1N1-S-OIV To create KBMA E. coli, the E. coli BL21(DE3) over-expressing HA of SD/H1N1-S-OIV or GFP was inactivated by UV irradiation at total energy of 7000 J/m 2 by a Spectrolinker (Spectronics, Westbury, NY) ( 23 , 25 , 32 ).
Virus:Article Title: Vaccination with Killed but Metabolically Active E. coli Over-expressing Hemagglutinin Elicits Neutralizing Antibodies to H1N1 Swine Origin Influenza A Virus
Article Snippet: Subsequently, the inclusion bodies were refolded as recommended by the manufacture (Amersham Biosciences, Piscataway, NJ). .. To further confirm the protein expression, the lysates of E. coli , BL21(DE3) harboring SD/H1N1-S-OIV HA, GFP, or P. acnes CAMP factor were subjected to 10 % SDS-PAGE, immunoblotted on polyvinylidene fluoride (PVDF) membranes and incubated with sera from mice immunized with KBMA E. coli HA SD/H1N1-S-OIV or a commercial antibody to swine origin influenza A (H1N1) HA (ProSci Incorporated, Poway, CA, USA). fig ft0 fig mode=article f1 Figure 1 caption a4 caption a8 Estimation of the expression level of HA of H1N1-S-OIV in E. coli, BL21 (DE3) (1 × 10 8 CFU) Virus growth A clinical isolate of SD/H1N1-S-OIV and two seasonal influenza A/Brisbane/59/2007 (H1N1) virus (ATCCH1N1) and an A/Brisbane/10/2007 (H3N2)-like virus (ATCCH3N2) were grown in virus growth medium containing Dulbecco's modified Eagle's medium (DMEM) with glutamine, 0.2% bovine serum albumin (BSA), 25 mM Hepes and 2 μg/ml TPCK-trypsin (Sigma Chemical Co., St. Louis, Mo.) and propagated in Madin-Darby canine kidney (MDCK) cells as described in our previous publication ( 31 ). .. Intranasal immunization with KBMA E. coli HA SD/H1N1-S-OIV To create KBMA E. coli, the E. coli BL21(DE3) over-expressing HA of SD/H1N1-S-OIV or GFP was inactivated by UV irradiation at total energy of 7000 J/m 2 by a Spectrolinker (Spectronics, Westbury, NY) ( 23 , 25 , 32 ).
Modification:Article Title: Vaccination with Killed but Metabolically Active E. coli Over-expressing Hemagglutinin Elicits Neutralizing Antibodies to H1N1 Swine Origin Influenza A Virus
Article Snippet: Subsequently, the inclusion bodies were refolded as recommended by the manufacture (Amersham Biosciences, Piscataway, NJ). .. To further confirm the protein expression, the lysates of E. coli , BL21(DE3) harboring SD/H1N1-S-OIV HA, GFP, or P. acnes CAMP factor were subjected to 10 % SDS-PAGE, immunoblotted on polyvinylidene fluoride (PVDF) membranes and incubated with sera from mice immunized with KBMA E. coli HA SD/H1N1-S-OIV or a commercial antibody to swine origin influenza A (H1N1) HA (ProSci Incorporated, Poway, CA, USA). fig ft0 fig mode=article f1 Figure 1 caption a4 caption a8 Estimation of the expression level of HA of H1N1-S-OIV in E. coli, BL21 (DE3) (1 × 10 8 CFU) Virus growth A clinical isolate of SD/H1N1-S-OIV and two seasonal influenza A/Brisbane/59/2007 (H1N1) virus (ATCCH1N1) and an A/Brisbane/10/2007 (H3N2)-like virus (ATCCH3N2) were grown in virus growth medium containing Dulbecco's modified Eagle's medium (DMEM) with glutamine, 0.2% bovine serum albumin (BSA), 25 mM Hepes and 2 μg/ml TPCK-trypsin (Sigma Chemical Co., St. Louis, Mo.) and propagated in Madin-Darby canine kidney (MDCK) cells as described in our previous publication ( 31 ). .. Intranasal immunization with KBMA E. coli HA SD/H1N1-S-OIV To create KBMA E. coli, the E. coli BL21(DE3) over-expressing HA of SD/H1N1-S-OIV or GFP was inactivated by UV irradiation at total energy of 7000 J/m 2 by a Spectrolinker (Spectronics, Westbury, NY) ( 23 , 25 , 32 ).
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