Arginine as an Eluent Overcomes the Hindrance of Monoclonal Antibody Quantification by Dextran Sulfate in Protein A Affinity Chromatography

Kim Bong Gyun; Park Hong Woo

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Arginine as an Eluent Overcomes the Hindrance of Monoclonal Antibody Quantification by Dextran Sulfate in Protein A Affinity Chromatography

机译：精氨酸作为洗脱液克服了蛋白A亲和层析中硫酸右旋糖酐对单克隆抗体定量的阻碍

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Analytical chromatography using protein A affinity columns was employed for the fast and simple quantitative analysis of monoclonal antibodies (mAb) from suspension cultures of recombinant Chinese hamster ovary (rCHO) cells. Reliable results could not be obtained from analysis of rCHO cell culture supernatants containing dextran sulfate using elution buffers such as phosphate, glycine, or MgCl2. These problems increased as the number of analysis and the concentration of dextran sulfate in samples increased. Arginine was identified as an alternative eluent to overcome the hindrance by dextran sulfate. When the samples contain dextran sulfate up to 100 mg/L, the elution buffer containing 0.6-1.0 M arginine at pH 3.0-3.8 is useful for the effective analysis. Reproducible results in the mAb quantification could be obtained by this developed arginine elution buffer from rCHO cell culture supernatants containing dextran sulfate. (C) 2015 American Institute of Chemical Engineers

机译：使用蛋白A亲和柱进行分析色谱，可快速，简单地对重组中国仓鼠卵巢（rCHO）细胞悬浮培养物中的单克隆抗体（mAb）进行定量分析。使用洗脱缓冲液（例如磷酸盐，甘氨酸或MgCl2）分析含硫酸葡聚糖的rCHO细胞培养上清液无法获得可靠的结果。随着分析次数的增加和样品中硫酸葡聚糖浓度的增加，这些问题也随之增加。精氨酸被认为是克服硫酸葡聚糖的障碍的替代洗脱液。当样品中硫酸葡聚糖的含量高达100 mg / L时，在pH 3.0-3.8时含有0.6-1.0 M精氨酸的洗脱缓冲液可用于有效分析。通过这种开发的精氨酸洗脱缓冲液，可以从含有硫酸葡聚糖的rCHO细胞培养上清液中获得可重复的mAb定量结果。（C）2015美国化学工程师学会

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《Biotechnology Progress》 |2015年第6期|共6页
作者
Kim Bong Gyun; Park Hong Woo;
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正文语种 eng
中图分类生物科学;
关键词
antibody; arginine; dextran sulfate; protein A;

机译：抗体;精氨酸;硫酸葡聚糖;蛋白A;

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专利

1. Arginine as an Eluent Overcomes the Hindrance of Monoclonal Antibody Quantification by Dextran Sulfate in Protein A Affinity Chromatography [J] . Kim Bong Gyun, Park Hong Woo Biotechnology Progress . 2015,第6期

机译：精氨酸作为洗脱液克服了蛋白A亲和层析中硫酸右旋糖酐对单克隆抗体定量的阻碍
2. Arginine as an eluent for automated on-line Protein A/size exclusion chromatographic analysis of monoclonal antibody aggregates in cell culture [J] . Sean Wang, Anil Raghani Journal of chromatography, B. Analytical technologies in the biomedical and life sciences . 2014,第Null期

机译：精氨酸作为洗脱液，用于细胞培养中单克隆抗体聚集体的自动化在线蛋白质A /大小排阻色谱分析
3. Toll-like receptor 2 monoclonal antibody or/and Toll-like receptor 4 monoclonal antibody increase counts of Lactobacilli and Bifidobacteria in dextran sulfate sodium-induced colitis in mice. [J] . Dong L, Li J, Liu Y, Journal of gastroenterology and hepatology . 2012,第1期

机译：Toll样受体2单克隆抗体或/和Toll样受体4单克隆抗体增加了硫酸葡聚糖钠诱导的小鼠结肠炎中乳酸杆菌和双歧杆菌的计数。
4. Using Hydrophilic Interaction Chromatography - Mass Spectrometry for Heightened Product Characterization to Overcome Challenges with Hydrophobic Antibody Drug Conjugates and Monoclonal Antibodies [C] . Jacquelynn Smith, Matthew A. Lauber, Stephan M. Koza, ASMS Conference on Mass Spectrometry and Allied Topics . 2016

机译：使用亲水性相互作用色谱 - 质谱法加热的产品特征，以克服疏水性抗体药物缀合物和单克隆抗体的挑战
5. 1. Selective facilitated transport of biological compounds through a phospholipid bilayer using boronic acid complexation. 2. Influence of eluent anions in boronate affinity chromatography. 3. Photoregulation of enzyme activity: Synthesis and study of pho [D] . Westmark, Pamela Rae. 1996

机译：1.使用硼酸络合选择性促进生物化合物通过磷脂双层的运输。 2.洗脱液阴离子在硼酸酯亲和色谱中的影响。 3.酶活性的光调节：磷酸的合成与研究
6. Purification of Shiga toxin and Shiga-like toxins I and II by receptor analog affinity chromatography with immobilized P1 glycoprotein and production of cross-reactive monoclonal antibodies. [O] . A Donohue-Rolfe, D W Acheson, A V Kane, 1989

机译：通过固定有P1糖蛋白的受体类似物亲和层析纯化志贺毒素和志贺样毒素I和II并产生交叉反应性单克隆抗体。
7. Tandem Immobilized Metal-Ion Affinity Chromatography/ Immunoaffinity Purification of His-tagged Proteins - Evaluation of Two Anti-His-Tag Monoclonal Antibodies [O] . Kristian M. Müller, Katja M. Arndt, Konrad Bauer, 2007

机译：串联固定的金属离子亲和层析/免疫亲和纯化的他带标签的蛋白质-两种抗His标签单克隆抗体的评估
8. Calculation of Monoclonal Antibody Affinity Constants Directly from Antibody Dilution Curves [R] . Griswold, W. R., Nelson, D. P. 1985

机译：直接从抗体稀释曲线计算单克隆抗体亲和力常数

Arginine as an Eluent Overcomes the Hindrance of Monoclonal Antibody Quantification by Dextran Sulfate in Protein A Affinity Chromatography

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