chok1sv  (Lonza)


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    Name:
    CHOK1SV
    Description:

    Catalog Number:
    CHOK1SV
    Price:
    None
    Category:
    Cell line
    Applications:
    CHO Cell line construction, cell line development, cell line engineering, cell line selection, gene expression, protein production, stable transfection, transient transfection, cloning, recombinant protein production, electroporation, transfection, transfectant pools, stable pools, mini pools, mini-pools, clonal cell line, fed batch culture, mini bioreactor, bioreactor, AMBR, single use bioreactor, deep well plate, deep-well plates, monoclonal antibodies, bi-specific antibodies, Fc fusion proteins, minibodies, fusion proteins, recombinant proteins, difficult to express proteins, complex proteins,
    Cell Type:
    suspension adapted, CDACF adapted
    Cell Line:
    CHO
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    Structured Review

    Lonza chok1sv
    CHOK1SV

    https://www.bioz.com/result/chok1sv/product/Lonza
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    chok1sv - by Bioz Stars, 2021-09
    99/100 stars

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    Related Articles

    Over Expression:

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Transient Overexpression of NDPK-A With EBNA-1 Mediated Extrachromosomal Maintenance Further Enhances Recombinant Protein Expression We next looked to combine the nuclear shuttling protein expression of NDPK-A with the EBNA-1 based technology into CHOK1SV cells to determine if this could deliver transient expression beyond the capacity of either component individually. ..

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed. ..

    Recombinant:

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Transient Overexpression of NDPK-A With EBNA-1 Mediated Extrachromosomal Maintenance Further Enhances Recombinant Protein Expression We next looked to combine the nuclear shuttling protein expression of NDPK-A with the EBNA-1 based technology into CHOK1SV cells to determine if this could deliver transient expression beyond the capacity of either component individually. ..

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. We did not use vectors with the BS on the recombinant (eGFP) vector as the results in section “Assessment of Transient Expression of the Genes for NDPK-A and Chx10 and Complementary Consensus Binding Sequences in CHOK1SV Cells” suggested this did not improve expression over using the NDPK-A gene alone. ..

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed. ..

    Expressing:

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Transient Overexpression of NDPK-A With EBNA-1 Mediated Extrachromosomal Maintenance Further Enhances Recombinant Protein Expression We next looked to combine the nuclear shuttling protein expression of NDPK-A with the EBNA-1 based technology into CHOK1SV cells to determine if this could deliver transient expression beyond the capacity of either component individually. ..

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. We did not use vectors with the BS on the recombinant (eGFP) vector as the results in section “Assessment of Transient Expression of the Genes for NDPK-A and Chx10 and Complementary Consensus Binding Sequences in CHOK1SV Cells” suggested this did not improve expression over using the NDPK-A gene alone. ..

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Assessment of Transient Expression of the Genes for NDPK-A and Chx10 and Complementary Consensus Binding Sequences in CHOK1SV Cells Expression vectors encoding eGFP in addition to either NDPK-A or Chx10 genes that contain an NLS, the consensus binding sequence (BS) or a combination of the two were transfected into Lonza’s CHOK1SV cells and analysed by flow cytometry ( ). ..

    Article Title: An engineered tetra-valent antibody fully activates the Tie2 receptor with comparable potency to its natural ligand angiopoietin-1
    Article Snippet: .. To express the antibodies, expression vectors were transfected into FreeStyle 293 cells (Thermo Fisher Scientific), Expi 293 cells (Thermo Fisher Scientific) or CHO-K1SV cells (Lonza) and cultured for 5–7 days. ..

    Transfection:

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. We therefore established the system here by undertaking transfections with Lonza CHOK1SV cells using pDNA containing either the truncated EBNA-1 gene, the minimal oriP sequence, or a combination of the two on the same plasmid. ..

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Assessment of Transient Expression of the Genes for NDPK-A and Chx10 and Complementary Consensus Binding Sequences in CHOK1SV Cells Expression vectors encoding eGFP in addition to either NDPK-A or Chx10 genes that contain an NLS, the consensus binding sequence (BS) or a combination of the two were transfected into Lonza’s CHOK1SV cells and analysed by flow cytometry ( ). ..

    Article Title: An engineered tetra-valent antibody fully activates the Tie2 receptor with comparable potency to its natural ligand angiopoietin-1
    Article Snippet: .. To express the antibodies, expression vectors were transfected into FreeStyle 293 cells (Thermo Fisher Scientific), Expi 293 cells (Thermo Fisher Scientific) or CHO-K1SV cells (Lonza) and cultured for 5–7 days. ..

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed. ..

    Sequencing:

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. We therefore established the system here by undertaking transfections with Lonza CHOK1SV cells using pDNA containing either the truncated EBNA-1 gene, the minimal oriP sequence, or a combination of the two on the same plasmid. ..

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Assessment of Transient Expression of the Genes for NDPK-A and Chx10 and Complementary Consensus Binding Sequences in CHOK1SV Cells Expression vectors encoding eGFP in addition to either NDPK-A or Chx10 genes that contain an NLS, the consensus binding sequence (BS) or a combination of the two were transfected into Lonza’s CHOK1SV cells and analysed by flow cytometry ( ). ..

    Plasmid Preparation:

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. We therefore established the system here by undertaking transfections with Lonza CHOK1SV cells using pDNA containing either the truncated EBNA-1 gene, the minimal oriP sequence, or a combination of the two on the same plasmid. ..

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Our data support this, but further, also suggests that a decrease in the rate of nuclear import of plasmid DNA may be linked to the reduced growth observed when the EBNA-1 gene is introduced in the Lonza CHOK1SV or CHOK1SV GS-KO cells either transiently or stably, respectively. ..

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. We did not use vectors with the BS on the recombinant (eGFP) vector as the results in section “Assessment of Transient Expression of the Genes for NDPK-A and Chx10 and Complementary Consensus Binding Sequences in CHOK1SV Cells” suggested this did not improve expression over using the NDPK-A gene alone. ..

    Stable Transfection:

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Our data support this, but further, also suggests that a decrease in the rate of nuclear import of plasmid DNA may be linked to the reduced growth observed when the EBNA-1 gene is introduced in the Lonza CHOK1SV or CHOK1SV GS-KO cells either transiently or stably, respectively. ..

    Binding Assay:

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. We did not use vectors with the BS on the recombinant (eGFP) vector as the results in section “Assessment of Transient Expression of the Genes for NDPK-A and Chx10 and Complementary Consensus Binding Sequences in CHOK1SV Cells” suggested this did not improve expression over using the NDPK-A gene alone. ..

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Assessment of Transient Expression of the Genes for NDPK-A and Chx10 and Complementary Consensus Binding Sequences in CHOK1SV Cells Expression vectors encoding eGFP in addition to either NDPK-A or Chx10 genes that contain an NLS, the consensus binding sequence (BS) or a combination of the two were transfected into Lonza’s CHOK1SV cells and analysed by flow cytometry ( ). ..

    Flow Cytometry:

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression
    Article Snippet: .. Assessment of Transient Expression of the Genes for NDPK-A and Chx10 and Complementary Consensus Binding Sequences in CHOK1SV Cells Expression vectors encoding eGFP in addition to either NDPK-A or Chx10 genes that contain an NLS, the consensus binding sequence (BS) or a combination of the two were transfected into Lonza’s CHOK1SV cells and analysed by flow cytometry ( ). ..

    Cell Culture:

    Article Title: An engineered tetra-valent antibody fully activates the Tie2 receptor with comparable potency to its natural ligand angiopoietin-1
    Article Snippet: .. To express the antibodies, expression vectors were transfected into FreeStyle 293 cells (Thermo Fisher Scientific), Expi 293 cells (Thermo Fisher Scientific) or CHO-K1SV cells (Lonza) and cultured for 5–7 days. ..

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    Lonza chok1sv gs ko cell pools
    Co-transfections were performed with Lonza <t>CHOK1SV</t> cells using different combinations of control (eGFP alone), NDPK-A NLS and EBNA1/ oriP vectors. X = control; ● = NDPK-A NLS and control; ◼ = EBNA-1/oriP and control, ▲ = EBNA-1/oriP and NDPK-A NLS (40 μg total DNA was used for each transfection, comprising 20 μg of each specified construct, except in the control samples where 40 μg of the control plasmid was transfected). Flow cytometry was used to assess the transient populations at different time points post transfection to ascertain (A) the percentage of cells exceeding a predetermined fluorescence threshold, and (B) the overall geometric mean fluorescence. Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).
    Chok1sv Gs Ko Cell Pools, supplied by Lonza, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/chok1sv gs ko cell pools/product/Lonza
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    chok1sv gs ko cell pools - by Bioz Stars, 2021-09
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    Co-transfections were performed with Lonza CHOK1SV cells using different combinations of control (eGFP alone), NDPK-A NLS and EBNA1/ oriP vectors. X = control; ● = NDPK-A NLS and control; ◼ = EBNA-1/oriP and control, ▲ = EBNA-1/oriP and NDPK-A NLS (40 μg total DNA was used for each transfection, comprising 20 μg of each specified construct, except in the control samples where 40 μg of the control plasmid was transfected). Flow cytometry was used to assess the transient populations at different time points post transfection to ascertain (A) the percentage of cells exceeding a predetermined fluorescence threshold, and (B) the overall geometric mean fluorescence. Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression

    doi: 10.3389/fbioe.2021.679448

    Figure Lengend Snippet: Co-transfections were performed with Lonza CHOK1SV cells using different combinations of control (eGFP alone), NDPK-A NLS and EBNA1/ oriP vectors. X = control; ● = NDPK-A NLS and control; ◼ = EBNA-1/oriP and control, ▲ = EBNA-1/oriP and NDPK-A NLS (40 μg total DNA was used for each transfection, comprising 20 μg of each specified construct, except in the control samples where 40 μg of the control plasmid was transfected). Flow cytometry was used to assess the transient populations at different time points post transfection to ascertain (A) the percentage of cells exceeding a predetermined fluorescence threshold, and (B) the overall geometric mean fluorescence. Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Article Snippet: Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed.

    Techniques: Transfection, Construct, Plasmid Preparation, Flow Cytometry, Fluorescence, Standard Deviation

    Analysis of Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with vectors containing genes for a model IgG mAb. (A) Shows western blot analysis of transient cultures transfected with a vector containing mAb genes alone or a mAb /oriP vector (denoted by “oriP ” on the figure). (B) Shows relative eGFP abundance from lysates of transient cultures (described above) harvested at 192 hours post-transfection. Quantification of IgG mAb was carried out using Octet analysis. A one way ANOVA test followed by post hoc Tukey test was carried out on titre data (B,C) . Conditions which do not share a letter are statistically different at the 95% confidence level. Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression

    doi: 10.3389/fbioe.2021.679448

    Figure Lengend Snippet: Analysis of Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with vectors containing genes for a model IgG mAb. (A) Shows western blot analysis of transient cultures transfected with a vector containing mAb genes alone or a mAb /oriP vector (denoted by “oriP ” on the figure). (B) Shows relative eGFP abundance from lysates of transient cultures (described above) harvested at 192 hours post-transfection. Quantification of IgG mAb was carried out using Octet analysis. A one way ANOVA test followed by post hoc Tukey test was carried out on titre data (B,C) . Conditions which do not share a letter are statistically different at the 95% confidence level. Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Article Snippet: Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed.

    Techniques: Generated, Construct, Transfection, Western Blot, Plasmid Preparation, Standard Deviation

    Analysis of Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with vectors containing the gene for a SARS nCov Spike and the oriP elements. (A) Shows western blot analysis of supernatants harvested 8 days post-transfection using an anti-His tag antibody, whilst figure (B) shows relative band intensities as determined using densitometry. A one way ANOVA test followed by post hoc Tukey test was carried out on densitometry data. Conditions which do not share a letter are statistically different at the 95% confidence level. Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression

    doi: 10.3389/fbioe.2021.679448

    Figure Lengend Snippet: Analysis of Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with vectors containing the gene for a SARS nCov Spike and the oriP elements. (A) Shows western blot analysis of supernatants harvested 8 days post-transfection using an anti-His tag antibody, whilst figure (B) shows relative band intensities as determined using densitometry. A one way ANOVA test followed by post hoc Tukey test was carried out on densitometry data. Conditions which do not share a letter are statistically different at the 95% confidence level. Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Article Snippet: Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed.

    Techniques: Generated, Construct, Transfection, Western Blot, Standard Deviation

    Vectors containing a truncated EBNA-1 gene, complementary oriP elements or a combination of the two were transiently electroporated in Lonza CHOK1SV cells and analysed every 24 hours post transfection for 216 hours. X = control (eGFP only), ◼ = oriP, ▲ = EBNA-1, ● = EBNA-1/oriP. (A) Viable cell number of CHOK1SV cells post transfection with the described constructs, (B) flow cytometry obtained values showing the percentage of cells of transient populations exceeding a predetermined fluorescence threshold, and (C) the overall mean fluorescence, were recorded. Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression

    doi: 10.3389/fbioe.2021.679448

    Figure Lengend Snippet: Vectors containing a truncated EBNA-1 gene, complementary oriP elements or a combination of the two were transiently electroporated in Lonza CHOK1SV cells and analysed every 24 hours post transfection for 216 hours. X = control (eGFP only), ◼ = oriP, ▲ = EBNA-1, ● = EBNA-1/oriP. (A) Viable cell number of CHOK1SV cells post transfection with the described constructs, (B) flow cytometry obtained values showing the percentage of cells of transient populations exceeding a predetermined fluorescence threshold, and (C) the overall mean fluorescence, were recorded. Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Article Snippet: Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed.

    Techniques: Transfection, Construct, Flow Cytometry, Fluorescence, Standard Deviation

    Flow cytometry analysis of Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with eGFP containing vectors. (A) Shows the percentage of cells exceeding a fluorescent intensity threshold in transient cultures transfected with a vector containing eGFP alone (X = Control, ○ = NDPK-A, □ = EBNA1 and △ = NDPK-A and EBNA1 cell pools). (B) Shows the percentage of cells exceeding a fluorescent intensity threshold in transient cultures transfected with a vector containing eGFP and oriP (X = Control, ● = NDPK-A, ◼ = EBNA1 and ▲ = NDPK-A and EBNA1 cell pools). (C) Shows the overall geometric mean fluorescence of transient cultures transfected with a vector containing eGFP alone (X = Control, ○ = NDPK-A, □ = EBNA1 and △ = NDPK-A EBNA1 cell pools). (D) Shows the overall geometric mean fluorescence of transient cultures transfected with a vector containing eGFP and oriP (X = Control, ● = NDPK-A, ◼ = EBNA1 and ▲ = NDPK-A and EBNA1 cell pools). Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression

    doi: 10.3389/fbioe.2021.679448

    Figure Lengend Snippet: Flow cytometry analysis of Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with eGFP containing vectors. (A) Shows the percentage of cells exceeding a fluorescent intensity threshold in transient cultures transfected with a vector containing eGFP alone (X = Control, ○ = NDPK-A, □ = EBNA1 and △ = NDPK-A and EBNA1 cell pools). (B) Shows the percentage of cells exceeding a fluorescent intensity threshold in transient cultures transfected with a vector containing eGFP and oriP (X = Control, ● = NDPK-A, ◼ = EBNA1 and ▲ = NDPK-A and EBNA1 cell pools). (C) Shows the overall geometric mean fluorescence of transient cultures transfected with a vector containing eGFP alone (X = Control, ○ = NDPK-A, □ = EBNA1 and △ = NDPK-A EBNA1 cell pools). (D) Shows the overall geometric mean fluorescence of transient cultures transfected with a vector containing eGFP and oriP (X = Control, ● = NDPK-A, ◼ = EBNA1 and ▲ = NDPK-A and EBNA1 cell pools). Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Article Snippet: Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed.

    Techniques: Flow Cytometry, Generated, Construct, Transfection, Plasmid Preparation, Fluorescence, Standard Deviation

    Viable cell number in Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with eGFP containing vectors. (A) Shows viable cell numbers of transient cultures transfected with a vector containing eGFP alone (X = Control, ○ = NDPK-A, □ = EBNA1 and △ = NDPK-A EBNA1 cell pools). (B) Shows viable cell numbers of transient cultures transfected with a vector containing eGFP and oriP elements (X = Control, ● = NDPK-A, ◼ = EBNA1 and ▲ = NDPK-A and EBNA1 cell pools). Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression

    doi: 10.3389/fbioe.2021.679448

    Figure Lengend Snippet: Viable cell number in Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with eGFP containing vectors. (A) Shows viable cell numbers of transient cultures transfected with a vector containing eGFP alone (X = Control, ○ = NDPK-A, □ = EBNA1 and △ = NDPK-A EBNA1 cell pools). (B) Shows viable cell numbers of transient cultures transfected with a vector containing eGFP and oriP elements (X = Control, ● = NDPK-A, ◼ = EBNA1 and ▲ = NDPK-A and EBNA1 cell pools). Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Article Snippet: Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed.

    Techniques: Generated, Construct, Transfection, Plasmid Preparation, Standard Deviation

    Analysis of Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with eGFP containing vectors. (A) Shows western blot analysis of transient cultures transfected with a vector containing eGFP alone or an eGFP /oriP vector (denoted by “oriP ” on the figure). (B) Shows relative eGFP abundance from lysates of transient cultures (described above) harvested at 192 h post-transfection. Densitometry analysis was carried out using ImageJ on western blots run in biological triplicate where β-actin was used as a loading control for normalisation. A one way ANOVA test followed by post hoc Tukey test was carried out on densitometry data. Conditions which do not share a letter are statistically different at the 95% confidence level. Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression

    doi: 10.3389/fbioe.2021.679448

    Figure Lengend Snippet: Analysis of Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with eGFP containing vectors. (A) Shows western blot analysis of transient cultures transfected with a vector containing eGFP alone or an eGFP /oriP vector (denoted by “oriP ” on the figure). (B) Shows relative eGFP abundance from lysates of transient cultures (described above) harvested at 192 h post-transfection. Densitometry analysis was carried out using ImageJ on western blots run in biological triplicate where β-actin was used as a loading control for normalisation. A one way ANOVA test followed by post hoc Tukey test was carried out on densitometry data. Conditions which do not share a letter are statistically different at the 95% confidence level. Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Article Snippet: Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed.

    Techniques: Generated, Construct, Transfection, Western Blot, Plasmid Preparation, Standard Deviation

    Vectors containing NDPK-A and Chx10 genes that include NLSs, complementary BSs or a combination of the two (vector components present on each vector are indicated through ± below corresponding bars) were transiently electroporated into Lonza CHOK1SV cells and the subsequent fluorescence intensity analysed at 24 and 48 h post transfection via flow cytometry. (A) The percentage of cells in the resulting transient populations exceeding a predetermined fluorescence intensity threshold, and (B) the overall geometric mean fluorescence. All experiments were run in biological triplicate ( n = 3) and conditions significantly different from the control condition for each time point are indicated on the graph ( p

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression

    doi: 10.3389/fbioe.2021.679448

    Figure Lengend Snippet: Vectors containing NDPK-A and Chx10 genes that include NLSs, complementary BSs or a combination of the two (vector components present on each vector are indicated through ± below corresponding bars) were transiently electroporated into Lonza CHOK1SV cells and the subsequent fluorescence intensity analysed at 24 and 48 h post transfection via flow cytometry. (A) The percentage of cells in the resulting transient populations exceeding a predetermined fluorescence intensity threshold, and (B) the overall geometric mean fluorescence. All experiments were run in biological triplicate ( n = 3) and conditions significantly different from the control condition for each time point are indicated on the graph ( p

    Article Snippet: Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed.

    Techniques: Plasmid Preparation, Fluorescence, Transfection, Flow Cytometry

    Viable cell number of Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with vectors containing genes for a model IgG4 mAb. (A) Shows viable cell numbers of transient cultures transfected with a vector containing the mAb genes alone (X = Control, ○ = NDPK-A, □ = EBNA1 and △ = NDPK-A and EBNA1 cell pools). (B) Shows viable cell numbers of transient cultures transfected with a vector containing mAb genes and oriP elements (X = Control, ● = NDPK-A, ◼ = EBNA1 and ▲ = NDPK-A and EBNA1 cell pools). Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Engineering of Chinese Hamster Ovary Cells With NDPK-A to Enhance DNA Nuclear Delivery Combined With EBNA1 Plasmid Maintenance Gives Improved Exogenous Transient Reporter, mAb and SARS-CoV-2 Spike Protein Expression

    doi: 10.3389/fbioe.2021.679448

    Figure Lengend Snippet: Viable cell number of Lonza CHOK1SV GS-KO cells engineered to overexpress NDPK-A, express EBNA-1 or both (control pools were generated using an empty construct) post-transfection with vectors containing genes for a model IgG4 mAb. (A) Shows viable cell numbers of transient cultures transfected with a vector containing the mAb genes alone (X = Control, ○ = NDPK-A, □ = EBNA1 and △ = NDPK-A and EBNA1 cell pools). (B) Shows viable cell numbers of transient cultures transfected with a vector containing mAb genes and oriP elements (X = Control, ● = NDPK-A, ◼ = EBNA1 and ▲ = NDPK-A and EBNA1 cell pools). Error bars show ± one standard deviation. All experiments were carried out in biological triplicate ( n = 3).

    Article Snippet: Overexpression of the NDPK-A gene in the Lonza CHOK1SV GS-KO cell pools resulted in enhanced recombinant eGFP yields upon transient transfection with both the eGFP or eGFP/oriP vectors that was evident by the increased band intensities observed.

    Techniques: Generated, Construct, Transfection, Plasmid Preparation, Standard Deviation

    ( a ) Western Blot of supernatants of transfected HEK293T cells expressing VEGFR1(D1-D3)-Fc protein at different time points in 24-well plate; ( b ) Western Blot for VEGFR1-Fc detection in supernatants of transfected CHOK1SV GS-KO clones at 24-well plate stage of clone development; ( c ) Western Blot of supernatants of clones at T-25 stage for VEGFR1(D1–D3)-Fc protein expression analysis; ( d ) Western Blot of supernatants of clones at T-75 stage for VEGFR1(D1–D3)-Fc protein expression analysis.

    Journal: International Journal of Molecular Sciences

    Article Title: Studies to Prevent Degradation of Recombinant Fc-Fusion Protein Expressed in Mammalian Cell Line and Protein Characterization

    doi: 10.3390/ijms17060913

    Figure Lengend Snippet: ( a ) Western Blot of supernatants of transfected HEK293T cells expressing VEGFR1(D1-D3)-Fc protein at different time points in 24-well plate; ( b ) Western Blot for VEGFR1-Fc detection in supernatants of transfected CHOK1SV GS-KO clones at 24-well plate stage of clone development; ( c ) Western Blot of supernatants of clones at T-25 stage for VEGFR1(D1–D3)-Fc protein expression analysis; ( d ) Western Blot of supernatants of clones at T-75 stage for VEGFR1(D1–D3)-Fc protein expression analysis.

    Article Snippet: Cell Culture CHOK1SV GS-KO (Lonza Biologics, Slough, Berkshire, UK) was the mammalian cell line used for producing VEGFR1(D1–D3)-Fc protein in culture.

    Techniques: Western Blot, Transfection, Expressing, Clone Assay

    Analysis of SREBF1 engineered CHOK1SV GS-KO™ host cells stably expressing a variety of secretory recombinant biotherapeutic molecules. Cell pools were constructed by the simultaneous co-transfection of two expression vectors. The first vector contained the GS gene for selection as well as genes appropriate for production of either cB72.3, FcFP or DTE-IgG1 whilst the second vector contained the SREBF1 gene driven by different promoters to yield either low (SREBF1 Low) or high (SREBF1 High) SREBF1 protein abundance. Control pools (Control) were generated using the appropriate first vector (also generated using GS selection) in conjunction with an empty second vector (i.e. not encoding the SREBF1 gene). Western blot analysis was carried out to determine relative levels of SREBF1 abundance from cell pools constructed to express either cB72.3 (A), FcFP (B) or DTE-IgG1 (C) where SREBF1 Low, SREBF1 High or a Control vector were co-transfected during the generation of pools as highlighted in the figure (L7a was used as a loading control). Figure D shows bands from a Coomassie blue stained SDS-PAGE gel and densitometry analysis of bands corresponding to the recombinant products harvested at 7 days of batch culture from the constructed cell pools. All lanes show bands corresponding to an independent cell pool construction.

    Journal: Metabolic Engineering

    Article Title: Engineering of Chinese hamster ovary cell lipid metabolism results in an expanded ER and enhanced recombinant biotherapeutic protein production

    doi: 10.1016/j.ymben.2019.11.007

    Figure Lengend Snippet: Analysis of SREBF1 engineered CHOK1SV GS-KO™ host cells stably expressing a variety of secretory recombinant biotherapeutic molecules. Cell pools were constructed by the simultaneous co-transfection of two expression vectors. The first vector contained the GS gene for selection as well as genes appropriate for production of either cB72.3, FcFP or DTE-IgG1 whilst the second vector contained the SREBF1 gene driven by different promoters to yield either low (SREBF1 Low) or high (SREBF1 High) SREBF1 protein abundance. Control pools (Control) were generated using the appropriate first vector (also generated using GS selection) in conjunction with an empty second vector (i.e. not encoding the SREBF1 gene). Western blot analysis was carried out to determine relative levels of SREBF1 abundance from cell pools constructed to express either cB72.3 (A), FcFP (B) or DTE-IgG1 (C) where SREBF1 Low, SREBF1 High or a Control vector were co-transfected during the generation of pools as highlighted in the figure (L7a was used as a loading control). Figure D shows bands from a Coomassie blue stained SDS-PAGE gel and densitometry analysis of bands corresponding to the recombinant products harvested at 7 days of batch culture from the constructed cell pools. All lanes show bands corresponding to an independent cell pool construction.

    Article Snippet: In this study we show that overexpression of SCD1 and SREBF1 in the Lonza Biologics’ proprietary CHOK1SV GS-KO™ host cell line; a glutamine synthetase (GS) knockout host cell line, results in enhanced secreted recombinant protein yields of a number of different model biotherapeutic proteins, although this is dependent upon the amount of overexpression of the lipid metabolism modifying (LMM) genes (SCD1 and SREBF1).

    Techniques: Stable Transfection, Expressing, Recombinant, Construct, Cotransfection, Plasmid Preparation, Selection, Generated, Western Blot, Transfection, Staining, SDS Page

    Analysis of cellular lipid content in control and LMM engineered CHO cells using mass spectrometry. (A) and (B) show Principal Component Analysis (PCA) of the mass spectrometry derived lipid profiles extracted from cells overexpressing either SREBF1 or SCD1 genes highlighting differences in lipidomic profiles of engineered CHOK1SV GS-KO cell lines harvested after 6 days of batch culture. Figure A shows PCA of control samples and samples engineered to overexpress SCD1; black data points represent controls (CHO-Ctl-1 = ●, CHO-Ctl-2 = ▲, CHO-Ctl-3 = +, CHO-Control POOL = ■) whilst blue data points represent SCD1 overexpressing cells (CHO-SCD1 LOW = Image 1 , CHO-SCD1 MID = +, CHO-SCD1 HIGH = Image 2 , CHO-SCD1 POOL = Image 3 ). Figure B shows analysis of control samples and samples engineered to overexpress SREBF1; black data points represent controls (CHO-Ctl-1 = ●, CHO-Ctl-2 = ▲, CHO-Ctl-3 = +, CHO-Control POOL = ■) whilst red data points represent SREBF1 overexpressing cells (CHO-SREBF1 LOW = Image 4 , CHO-SREBF1 MID1 = Image 5 , CHO-SREBF1 MID2 = Image 6 , CHO-SREBF1 POOL = Image 7 ). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

    Journal: Metabolic Engineering

    Article Title: Engineering of Chinese hamster ovary cell lipid metabolism results in an expanded ER and enhanced recombinant biotherapeutic protein production

    doi: 10.1016/j.ymben.2019.11.007

    Figure Lengend Snippet: Analysis of cellular lipid content in control and LMM engineered CHO cells using mass spectrometry. (A) and (B) show Principal Component Analysis (PCA) of the mass spectrometry derived lipid profiles extracted from cells overexpressing either SREBF1 or SCD1 genes highlighting differences in lipidomic profiles of engineered CHOK1SV GS-KO cell lines harvested after 6 days of batch culture. Figure A shows PCA of control samples and samples engineered to overexpress SCD1; black data points represent controls (CHO-Ctl-1 = ●, CHO-Ctl-2 = ▲, CHO-Ctl-3 = +, CHO-Control POOL = ■) whilst blue data points represent SCD1 overexpressing cells (CHO-SCD1 LOW = Image 1 , CHO-SCD1 MID = +, CHO-SCD1 HIGH = Image 2 , CHO-SCD1 POOL = Image 3 ). Figure B shows analysis of control samples and samples engineered to overexpress SREBF1; black data points represent controls (CHO-Ctl-1 = ●, CHO-Ctl-2 = ▲, CHO-Ctl-3 = +, CHO-Control POOL = ■) whilst red data points represent SREBF1 overexpressing cells (CHO-SREBF1 LOW = Image 4 , CHO-SREBF1 MID1 = Image 5 , CHO-SREBF1 MID2 = Image 6 , CHO-SREBF1 POOL = Image 7 ). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

    Article Snippet: In this study we show that overexpression of SCD1 and SREBF1 in the Lonza Biologics’ proprietary CHOK1SV GS-KO™ host cell line; a glutamine synthetase (GS) knockout host cell line, results in enhanced secreted recombinant protein yields of a number of different model biotherapeutic proteins, although this is dependent upon the amount of overexpression of the lipid metabolism modifying (LMM) genes (SCD1 and SREBF1).

    Techniques: Mass Spectrometry, Derivative Assay

    Overexpression and cellular localization of SCD1 and SREBF1 in engineered cells. Western blot analysis of CHOK1SV GS-KO™ cells engineered to overexpress either SCD1 or SREBF1 genes where controls have been generated with an unmodified pcDNA3.1V5-His/TOPO vector. CHO-Ctl-1, CHO-Ctl-2 and CHO-Ctl-3 are monoclonal lines derived from the CHO-Control POOL ; CHO-SCD1 LOW , CHO-SCD1 MID and CHO-SCD1 HIGH are monoclonal cell lines isolated from CHO-SCD1 POOL and CHO-SREBF1 LOW , CHO-SREBF1 MID1 and CHO-SREBF1 MID2 are monoclonal lines isolated from CHO-SREBF1 POOL . The pools which had been engineered were used to generate cells expressing cB72.3 (CHO-Control cB72.3 , CHO-SCD1 cB72.3 and CHO-SREBF1 cB72.3 ) or an Fc fusion protein (FcFP; CHO-Control FcFP , CHO-SCD1 FcFP and CHO-SREBF1 FcFP ). Lysate samples were generated from these cells harvested at day 6 of culture from either host cells (A) or cells stably producing the recombinant products cB72.3 or FcFP where specified (B). A primary antibody with specificity for L7α was used as a loading control. (C) Cellular localization of overexpressed SCD1 and SREBF1 proteins in CHO-Control POOL , CHO-SCD1 POOL and CHO-SREBF1 POOL cell pools as determined by immunofluorescent detection using an anti-V5 antibody conjugated with a FITC secondary antibody. An anti-calnexin antibody conjugated with a TRITC secondary antibody was used to highlight the position of the ER. Images were obtained using confocal microscopy (see Data in Brief ( Budge et al., 2019 ) Table 2 for full details of antibodies used).

    Journal: Metabolic Engineering

    Article Title: Engineering of Chinese hamster ovary cell lipid metabolism results in an expanded ER and enhanced recombinant biotherapeutic protein production

    doi: 10.1016/j.ymben.2019.11.007

    Figure Lengend Snippet: Overexpression and cellular localization of SCD1 and SREBF1 in engineered cells. Western blot analysis of CHOK1SV GS-KO™ cells engineered to overexpress either SCD1 or SREBF1 genes where controls have been generated with an unmodified pcDNA3.1V5-His/TOPO vector. CHO-Ctl-1, CHO-Ctl-2 and CHO-Ctl-3 are monoclonal lines derived from the CHO-Control POOL ; CHO-SCD1 LOW , CHO-SCD1 MID and CHO-SCD1 HIGH are monoclonal cell lines isolated from CHO-SCD1 POOL and CHO-SREBF1 LOW , CHO-SREBF1 MID1 and CHO-SREBF1 MID2 are monoclonal lines isolated from CHO-SREBF1 POOL . The pools which had been engineered were used to generate cells expressing cB72.3 (CHO-Control cB72.3 , CHO-SCD1 cB72.3 and CHO-SREBF1 cB72.3 ) or an Fc fusion protein (FcFP; CHO-Control FcFP , CHO-SCD1 FcFP and CHO-SREBF1 FcFP ). Lysate samples were generated from these cells harvested at day 6 of culture from either host cells (A) or cells stably producing the recombinant products cB72.3 or FcFP where specified (B). A primary antibody with specificity for L7α was used as a loading control. (C) Cellular localization of overexpressed SCD1 and SREBF1 proteins in CHO-Control POOL , CHO-SCD1 POOL and CHO-SREBF1 POOL cell pools as determined by immunofluorescent detection using an anti-V5 antibody conjugated with a FITC secondary antibody. An anti-calnexin antibody conjugated with a TRITC secondary antibody was used to highlight the position of the ER. Images were obtained using confocal microscopy (see Data in Brief ( Budge et al., 2019 ) Table 2 for full details of antibodies used).

    Article Snippet: In this study we show that overexpression of SCD1 and SREBF1 in the Lonza Biologics’ proprietary CHOK1SV GS-KO™ host cell line; a glutamine synthetase (GS) knockout host cell line, results in enhanced secreted recombinant protein yields of a number of different model biotherapeutic proteins, although this is dependent upon the amount of overexpression of the lipid metabolism modifying (LMM) genes (SCD1 and SREBF1).

    Techniques: Over Expression, Western Blot, Generated, Plasmid Preparation, Derivative Assay, Isolation, Expressing, Stable Transfection, Recombinant, Confocal Microscopy