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final cation exchange chromatography step  (Repligen Corp)


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    Repligen Corp final cation exchange chromatography step
    Figure 1. Outline of experimental set-up. In the first case study, mAb 1 samples from a 50 L bioreactor run were taken at six points during the cultivation. Samples were centrifuged, filtered and then protein A purified, before being quantified for HCPs. In the second study, a DoE with mAb 2 was carried out to explore two cell culture parameters in order to determine the optimal upstream process conditions for high titer and low HCP levels. In the third study, mAb 3 was grown in shake flasks under fixed upstream parameters and cell broth samples were taken at four time points. At each time point, half of the material was sheared (2) while the other half of the material was left non-sheared (1). Both materials were subsequently centrifuged, filtered, and protein A purified. Next, all protein A eluate samples were further processed by anion exchange <t>chromatography</t> in flow-through mode and by cation exchange chromatography. All samples were analysed for product quality by measuring monomer percentage and HCP levels. Figure 2. Effects of harvest time on product titer and HCP levels (Case Study I). Increasing product amount over the course of a cultivation is accompanied by decreasing product purity. Supernatant samples were taken throughout the 50 L STR production bioreactor run of mAb 1 (including on harvest day) and antibody concentration was measured and normalised to 1 due to confidentiality. Supernatant samples were filtered, protein A purified, and analysed for HCP quantities by ELISA. Note that the HCP data points for days 1, 3 and 7 are estimated on titer as HCP levels were below detection (< 2 ng/mL). Descriptive error bars are based on assay serial dilutions and show one standard deviation. Figure 3. Effects of culture conditions (Case Study II). (A) Experimental DoE points. 14 fed- batch cultures were grown in 250 mL small scale bioreactor vessels (TAP ambr250) under a range of different pHs and temperatures. The empty, half-filled and filled symbols represent low, medium and high temperatures respectively, whereas the squares, triangles and circles represent low, medium and high pH points. All cultures were harvested on day 16. (B) Harvested samples were centrifuged, filtered and protein A purified, in order to determine HCP quantities, and correlate HCP data to titer results. Descriptive error bars are based on HCP assay serial dilutions and show one standard deviation. (C) Peak cell densities and cell- specific productivity rates were determined for each sample. Highlighted in the grey oval are cultures associated with low titers.
    Final Cation Exchange Chromatography Step, supplied by Repligen Corp, used in various techniques. Bioz Stars score: 95/100, based on 272 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/final cation exchange chromatography step/product/Repligen Corp
    Average 95 stars, based on 272 article reviews
    final cation exchange chromatography step - by Bioz Stars, 2026-03
    95/100 stars

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    1) Product Images from "Identification of upstream culture conditions and harvest time parameters that affect host cell protein clearance."

    Article Title: Identification of upstream culture conditions and harvest time parameters that affect host cell protein clearance.

    Journal: Biotechnology progress

    doi: 10.1002/btpr.2805

    Figure 1. Outline of experimental set-up. In the first case study, mAb 1 samples from a 50 L bioreactor run were taken at six points during the cultivation. Samples were centrifuged, filtered and then protein A purified, before being quantified for HCPs. In the second study, a DoE with mAb 2 was carried out to explore two cell culture parameters in order to determine the optimal upstream process conditions for high titer and low HCP levels. In the third study, mAb 3 was grown in shake flasks under fixed upstream parameters and cell broth samples were taken at four time points. At each time point, half of the material was sheared (2) while the other half of the material was left non-sheared (1). Both materials were subsequently centrifuged, filtered, and protein A purified. Next, all protein A eluate samples were further processed by anion exchange chromatography in flow-through mode and by cation exchange chromatography. All samples were analysed for product quality by measuring monomer percentage and HCP levels. Figure 2. Effects of harvest time on product titer and HCP levels (Case Study I). Increasing product amount over the course of a cultivation is accompanied by decreasing product purity. Supernatant samples were taken throughout the 50 L STR production bioreactor run of mAb 1 (including on harvest day) and antibody concentration was measured and normalised to 1 due to confidentiality. Supernatant samples were filtered, protein A purified, and analysed for HCP quantities by ELISA. Note that the HCP data points for days 1, 3 and 7 are estimated on titer as HCP levels were below detection (< 2 ng/mL). Descriptive error bars are based on assay serial dilutions and show one standard deviation. Figure 3. Effects of culture conditions (Case Study II). (A) Experimental DoE points. 14 fed- batch cultures were grown in 250 mL small scale bioreactor vessels (TAP ambr250) under a range of different pHs and temperatures. The empty, half-filled and filled symbols represent low, medium and high temperatures respectively, whereas the squares, triangles and circles represent low, medium and high pH points. All cultures were harvested on day 16. (B) Harvested samples were centrifuged, filtered and protein A purified, in order to determine HCP quantities, and correlate HCP data to titer results. Descriptive error bars are based on HCP assay serial dilutions and show one standard deviation. (C) Peak cell densities and cell- specific productivity rates were determined for each sample. Highlighted in the grey oval are cultures associated with low titers.
    Figure Legend Snippet: Figure 1. Outline of experimental set-up. In the first case study, mAb 1 samples from a 50 L bioreactor run were taken at six points during the cultivation. Samples were centrifuged, filtered and then protein A purified, before being quantified for HCPs. In the second study, a DoE with mAb 2 was carried out to explore two cell culture parameters in order to determine the optimal upstream process conditions for high titer and low HCP levels. In the third study, mAb 3 was grown in shake flasks under fixed upstream parameters and cell broth samples were taken at four time points. At each time point, half of the material was sheared (2) while the other half of the material was left non-sheared (1). Both materials were subsequently centrifuged, filtered, and protein A purified. Next, all protein A eluate samples were further processed by anion exchange chromatography in flow-through mode and by cation exchange chromatography. All samples were analysed for product quality by measuring monomer percentage and HCP levels. Figure 2. Effects of harvest time on product titer and HCP levels (Case Study I). Increasing product amount over the course of a cultivation is accompanied by decreasing product purity. Supernatant samples were taken throughout the 50 L STR production bioreactor run of mAb 1 (including on harvest day) and antibody concentration was measured and normalised to 1 due to confidentiality. Supernatant samples were filtered, protein A purified, and analysed for HCP quantities by ELISA. Note that the HCP data points for days 1, 3 and 7 are estimated on titer as HCP levels were below detection (< 2 ng/mL). Descriptive error bars are based on assay serial dilutions and show one standard deviation. Figure 3. Effects of culture conditions (Case Study II). (A) Experimental DoE points. 14 fed- batch cultures were grown in 250 mL small scale bioreactor vessels (TAP ambr250) under a range of different pHs and temperatures. The empty, half-filled and filled symbols represent low, medium and high temperatures respectively, whereas the squares, triangles and circles represent low, medium and high pH points. All cultures were harvested on day 16. (B) Harvested samples were centrifuged, filtered and protein A purified, in order to determine HCP quantities, and correlate HCP data to titer results. Descriptive error bars are based on HCP assay serial dilutions and show one standard deviation. (C) Peak cell densities and cell- specific productivity rates were determined for each sample. Highlighted in the grey oval are cultures associated with low titers.

    Techniques Used: Purification, Cell Culture, Chromatography, Concentration Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Figure 7. Effects of shear damage on product quality in early stage cell culture samples (Case Study III). Material from day 10 was processed as described in the legend for figure 1 (non- sheared vs. sheared) and aggregate / fragment levels were determined by analytical SEC after protein A purification. Figure 8. Effects of a polishing chromatography step on product quality (Case Study III). (A) HCP levels in non-sheared samples before and after cation exchange chromatography, as determined by ELISA. HCP data point for day 10 after CEX is approximated based on titer as HCP levels were below detection (< 2 ng/mL). Descriptive error bars are based on HCP assay serial dilutions and show one standard deviation. (B) Fragmentation percentages in non- sheared and sheared samples before and after cation exchange chromatography, as determined by analytical SEC.
    Figure Legend Snippet: Figure 7. Effects of shear damage on product quality in early stage cell culture samples (Case Study III). Material from day 10 was processed as described in the legend for figure 1 (non- sheared vs. sheared) and aggregate / fragment levels were determined by analytical SEC after protein A purification. Figure 8. Effects of a polishing chromatography step on product quality (Case Study III). (A) HCP levels in non-sheared samples before and after cation exchange chromatography, as determined by ELISA. HCP data point for day 10 after CEX is approximated based on titer as HCP levels were below detection (< 2 ng/mL). Descriptive error bars are based on HCP assay serial dilutions and show one standard deviation. (B) Fragmentation percentages in non- sheared and sheared samples before and after cation exchange chromatography, as determined by analytical SEC.

    Techniques Used: Shear, Cell Culture, Purification, Chromatography, Enzyme-linked Immunosorbent Assay, Standard Deviation



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    Repligen Corp final cation exchange chromatography step
    Figure 1. Outline of experimental set-up. In the first case study, mAb 1 samples from a 50 L bioreactor run were taken at six points during the cultivation. Samples were centrifuged, filtered and then protein A purified, before being quantified for HCPs. In the second study, a DoE with mAb 2 was carried out to explore two cell culture parameters in order to determine the optimal upstream process conditions for high titer and low HCP levels. In the third study, mAb 3 was grown in shake flasks under fixed upstream parameters and cell broth samples were taken at four time points. At each time point, half of the material was sheared (2) while the other half of the material was left non-sheared (1). Both materials were subsequently centrifuged, filtered, and protein A purified. Next, all protein A eluate samples were further processed by anion exchange <t>chromatography</t> in flow-through mode and by cation exchange chromatography. All samples were analysed for product quality by measuring monomer percentage and HCP levels. Figure 2. Effects of harvest time on product titer and HCP levels (Case Study I). Increasing product amount over the course of a cultivation is accompanied by decreasing product purity. Supernatant samples were taken throughout the 50 L STR production bioreactor run of mAb 1 (including on harvest day) and antibody concentration was measured and normalised to 1 due to confidentiality. Supernatant samples were filtered, protein A purified, and analysed for HCP quantities by ELISA. Note that the HCP data points for days 1, 3 and 7 are estimated on titer as HCP levels were below detection (< 2 ng/mL). Descriptive error bars are based on assay serial dilutions and show one standard deviation. Figure 3. Effects of culture conditions (Case Study II). (A) Experimental DoE points. 14 fed- batch cultures were grown in 250 mL small scale bioreactor vessels (TAP ambr250) under a range of different pHs and temperatures. The empty, half-filled and filled symbols represent low, medium and high temperatures respectively, whereas the squares, triangles and circles represent low, medium and high pH points. All cultures were harvested on day 16. (B) Harvested samples were centrifuged, filtered and protein A purified, in order to determine HCP quantities, and correlate HCP data to titer results. Descriptive error bars are based on HCP assay serial dilutions and show one standard deviation. (C) Peak cell densities and cell- specific productivity rates were determined for each sample. Highlighted in the grey oval are cultures associated with low titers.
    Final Cation Exchange Chromatography Step, supplied by Repligen Corp, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/final cation exchange chromatography step/product/Repligen Corp
    Average 95 stars, based on 1 article reviews
    final cation exchange chromatography step - by Bioz Stars, 2026-03
    95/100 stars
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    Figure 1. Outline of experimental set-up. In the first case study, mAb 1 samples from a 50 L bioreactor run were taken at six points during the cultivation. Samples were centrifuged, filtered and then protein A purified, before being quantified for HCPs. In the second study, a DoE with mAb 2 was carried out to explore two cell culture parameters in order to determine the optimal upstream process conditions for high titer and low HCP levels. In the third study, mAb 3 was grown in shake flasks under fixed upstream parameters and cell broth samples were taken at four time points. At each time point, half of the material was sheared (2) while the other half of the material was left non-sheared (1). Both materials were subsequently centrifuged, filtered, and protein A purified. Next, all protein A eluate samples were further processed by anion exchange chromatography in flow-through mode and by cation exchange chromatography. All samples were analysed for product quality by measuring monomer percentage and HCP levels. Figure 2. Effects of harvest time on product titer and HCP levels (Case Study I). Increasing product amount over the course of a cultivation is accompanied by decreasing product purity. Supernatant samples were taken throughout the 50 L STR production bioreactor run of mAb 1 (including on harvest day) and antibody concentration was measured and normalised to 1 due to confidentiality. Supernatant samples were filtered, protein A purified, and analysed for HCP quantities by ELISA. Note that the HCP data points for days 1, 3 and 7 are estimated on titer as HCP levels were below detection (< 2 ng/mL). Descriptive error bars are based on assay serial dilutions and show one standard deviation. Figure 3. Effects of culture conditions (Case Study II). (A) Experimental DoE points. 14 fed- batch cultures were grown in 250 mL small scale bioreactor vessels (TAP ambr250) under a range of different pHs and temperatures. The empty, half-filled and filled symbols represent low, medium and high temperatures respectively, whereas the squares, triangles and circles represent low, medium and high pH points. All cultures were harvested on day 16. (B) Harvested samples were centrifuged, filtered and protein A purified, in order to determine HCP quantities, and correlate HCP data to titer results. Descriptive error bars are based on HCP assay serial dilutions and show one standard deviation. (C) Peak cell densities and cell- specific productivity rates were determined for each sample. Highlighted in the grey oval are cultures associated with low titers.

    Journal: Biotechnology progress

    Article Title: Identification of upstream culture conditions and harvest time parameters that affect host cell protein clearance.

    doi: 10.1002/btpr.2805

    Figure Lengend Snippet: Figure 1. Outline of experimental set-up. In the first case study, mAb 1 samples from a 50 L bioreactor run were taken at six points during the cultivation. Samples were centrifuged, filtered and then protein A purified, before being quantified for HCPs. In the second study, a DoE with mAb 2 was carried out to explore two cell culture parameters in order to determine the optimal upstream process conditions for high titer and low HCP levels. In the third study, mAb 3 was grown in shake flasks under fixed upstream parameters and cell broth samples were taken at four time points. At each time point, half of the material was sheared (2) while the other half of the material was left non-sheared (1). Both materials were subsequently centrifuged, filtered, and protein A purified. Next, all protein A eluate samples were further processed by anion exchange chromatography in flow-through mode and by cation exchange chromatography. All samples were analysed for product quality by measuring monomer percentage and HCP levels. Figure 2. Effects of harvest time on product titer and HCP levels (Case Study I). Increasing product amount over the course of a cultivation is accompanied by decreasing product purity. Supernatant samples were taken throughout the 50 L STR production bioreactor run of mAb 1 (including on harvest day) and antibody concentration was measured and normalised to 1 due to confidentiality. Supernatant samples were filtered, protein A purified, and analysed for HCP quantities by ELISA. Note that the HCP data points for days 1, 3 and 7 are estimated on titer as HCP levels were below detection (< 2 ng/mL). Descriptive error bars are based on assay serial dilutions and show one standard deviation. Figure 3. Effects of culture conditions (Case Study II). (A) Experimental DoE points. 14 fed- batch cultures were grown in 250 mL small scale bioreactor vessels (TAP ambr250) under a range of different pHs and temperatures. The empty, half-filled and filled symbols represent low, medium and high temperatures respectively, whereas the squares, triangles and circles represent low, medium and high pH points. All cultures were harvested on day 16. (B) Harvested samples were centrifuged, filtered and protein A purified, in order to determine HCP quantities, and correlate HCP data to titer results. Descriptive error bars are based on HCP assay serial dilutions and show one standard deviation. (C) Peak cell densities and cell- specific productivity rates were determined for each sample. Highlighted in the grey oval are cultures associated with low titers.

    Article Snippet: The flow-through material was collected and polished with a final cation exchange chromatography step, using 1 mL MiniChrom columns (Repligen, Waltham, MA) and a buffer with a pH below the pI of the protein.

    Techniques: Purification, Cell Culture, Chromatography, Concentration Assay, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Figure 7. Effects of shear damage on product quality in early stage cell culture samples (Case Study III). Material from day 10 was processed as described in the legend for figure 1 (non- sheared vs. sheared) and aggregate / fragment levels were determined by analytical SEC after protein A purification. Figure 8. Effects of a polishing chromatography step on product quality (Case Study III). (A) HCP levels in non-sheared samples before and after cation exchange chromatography, as determined by ELISA. HCP data point for day 10 after CEX is approximated based on titer as HCP levels were below detection (< 2 ng/mL). Descriptive error bars are based on HCP assay serial dilutions and show one standard deviation. (B) Fragmentation percentages in non- sheared and sheared samples before and after cation exchange chromatography, as determined by analytical SEC.

    Journal: Biotechnology progress

    Article Title: Identification of upstream culture conditions and harvest time parameters that affect host cell protein clearance.

    doi: 10.1002/btpr.2805

    Figure Lengend Snippet: Figure 7. Effects of shear damage on product quality in early stage cell culture samples (Case Study III). Material from day 10 was processed as described in the legend for figure 1 (non- sheared vs. sheared) and aggregate / fragment levels were determined by analytical SEC after protein A purification. Figure 8. Effects of a polishing chromatography step on product quality (Case Study III). (A) HCP levels in non-sheared samples before and after cation exchange chromatography, as determined by ELISA. HCP data point for day 10 after CEX is approximated based on titer as HCP levels were below detection (< 2 ng/mL). Descriptive error bars are based on HCP assay serial dilutions and show one standard deviation. (B) Fragmentation percentages in non- sheared and sheared samples before and after cation exchange chromatography, as determined by analytical SEC.

    Article Snippet: The flow-through material was collected and polished with a final cation exchange chromatography step, using 1 mL MiniChrom columns (Repligen, Waltham, MA) and a buffer with a pH below the pI of the protein.

    Techniques: Shear, Cell Culture, Purification, Chromatography, Enzyme-linked Immunosorbent Assay, Standard Deviation