The Experts below are selected from a list of 27 Experts worldwide ranked by ideXlab platform
James Lambropoulos - One of the best experts on this subject based on the ideXlab platform.
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A high‐throughput capillary isoelectric focusing immunoassay for fingerprinting protein sialylation
Biotechnology Progress, 2015Co-Authors: Lam Raga A. Markely, Lila Cheung, Shashi Prajapati, Iva Turyan, Ruth Frenkel, Zoran Sosic, Scott Estes, Young Jun Choi, Thomas Ryll, James LambropoulosAbstract:The serum Half-Life, Biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high-throughput method for semi-quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:235–241, 2016
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A high-throughput capillary isoelectric focusing immunoassay for fingerprinting protein sialylation.
Biotechnology Progress, 2015Co-Authors: Lam Raga A. Markely, Lila Cheung, Shashi Prajapati, Iva Turyan, Ruth Frenkel, Zoran Sosic, Scott Estes, Young Jun Choi, Thomas Ryll, James LambropoulosAbstract:The serum Half-Life, Biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high-throughput method for semi-quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry.
Lam Raga A. Markely - One of the best experts on this subject based on the ideXlab platform.
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A high‐throughput capillary isoelectric focusing immunoassay for fingerprinting protein sialylation
Biotechnology Progress, 2015Co-Authors: Lam Raga A. Markely, Lila Cheung, Shashi Prajapati, Iva Turyan, Ruth Frenkel, Zoran Sosic, Scott Estes, Young Jun Choi, Thomas Ryll, James LambropoulosAbstract:The serum Half-Life, Biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high-throughput method for semi-quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:235–241, 2016
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A high-throughput capillary isoelectric focusing immunoassay for fingerprinting protein sialylation.
Biotechnology Progress, 2015Co-Authors: Lam Raga A. Markely, Lila Cheung, Shashi Prajapati, Iva Turyan, Ruth Frenkel, Zoran Sosic, Scott Estes, Young Jun Choi, Thomas Ryll, James LambropoulosAbstract:The serum Half-Life, Biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high-throughput method for semi-quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry.
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A high-throughput method for quantification of glycoprotein sialylation.
Analytical biochemistry, 2010Co-Authors: Lam Raga A. Markely, Boon Tee Ong, Kong Meng Hoi, Gavin Teo, Daniel I.c. WangAbstract:Sialic acid can improve qualities of therapeutic glycoproteins such as circulatory Half-Life, Biological activity, and solubility. In production of therapeutic glycoproteins, a high-throughput method is required for process monitoring and optimization to ensure consistent and optimal sialic acid content. Current methods for quantifying sialic acid, however, require chromatographic separation that is time-consuming and cannot rapidly analyze many samples in parallel. Here we present a novel high-throughput method for quantifying glycoprotein sialylation. Using chemical reduction, enzymatic release of sialic acid, and chemical derivatization of the sialic acid, the method can accurately, rapidly (15 min), and specifically analyze many samples in parallel. It requires only 45 μl of sample and has a quantitation limit of 2 μM sialic acid. It has also been validated for monitoring sialylation of recombinant interferon gamma (IFN-γ) produced in Chinese hamster ovary (CHO) cell culture. This method is useful for various applications in upstream and downstream bioprocesses.
Lila Cheung - One of the best experts on this subject based on the ideXlab platform.
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A high‐throughput capillary isoelectric focusing immunoassay for fingerprinting protein sialylation
Biotechnology Progress, 2015Co-Authors: Lam Raga A. Markely, Lila Cheung, Shashi Prajapati, Iva Turyan, Ruth Frenkel, Zoran Sosic, Scott Estes, Young Jun Choi, Thomas Ryll, James LambropoulosAbstract:The serum Half-Life, Biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high-throughput method for semi-quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:235–241, 2016
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A high-throughput capillary isoelectric focusing immunoassay for fingerprinting protein sialylation.
Biotechnology Progress, 2015Co-Authors: Lam Raga A. Markely, Lila Cheung, Shashi Prajapati, Iva Turyan, Ruth Frenkel, Zoran Sosic, Scott Estes, Young Jun Choi, Thomas Ryll, James LambropoulosAbstract:The serum Half-Life, Biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high-throughput method for semi-quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry.
Shashi Prajapati - One of the best experts on this subject based on the ideXlab platform.
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A high‐throughput capillary isoelectric focusing immunoassay for fingerprinting protein sialylation
Biotechnology Progress, 2015Co-Authors: Lam Raga A. Markely, Lila Cheung, Shashi Prajapati, Iva Turyan, Ruth Frenkel, Zoran Sosic, Scott Estes, Young Jun Choi, Thomas Ryll, James LambropoulosAbstract:The serum Half-Life, Biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high-throughput method for semi-quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:235–241, 2016
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A high-throughput capillary isoelectric focusing immunoassay for fingerprinting protein sialylation.
Biotechnology Progress, 2015Co-Authors: Lam Raga A. Markely, Lila Cheung, Shashi Prajapati, Iva Turyan, Ruth Frenkel, Zoran Sosic, Scott Estes, Young Jun Choi, Thomas Ryll, James LambropoulosAbstract:The serum Half-Life, Biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high-throughput method for semi-quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry.
Iva Turyan - One of the best experts on this subject based on the ideXlab platform.
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A high‐throughput capillary isoelectric focusing immunoassay for fingerprinting protein sialylation
Biotechnology Progress, 2015Co-Authors: Lam Raga A. Markely, Lila Cheung, Shashi Prajapati, Iva Turyan, Ruth Frenkel, Zoran Sosic, Scott Estes, Young Jun Choi, Thomas Ryll, James LambropoulosAbstract:The serum Half-Life, Biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high-throughput method for semi-quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 32:235–241, 2016
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A high-throughput capillary isoelectric focusing immunoassay for fingerprinting protein sialylation.
Biotechnology Progress, 2015Co-Authors: Lam Raga A. Markely, Lila Cheung, Shashi Prajapati, Iva Turyan, Ruth Frenkel, Zoran Sosic, Scott Estes, Young Jun Choi, Thomas Ryll, James LambropoulosAbstract:The serum Half-Life, Biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high-throughput method for semi-quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry.
