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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초록

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.

키워드

antibodyargininedextran sulfateprotein ASERUM-FREE MEDIUMRECOMBINANT ANTIBODYRATIONAL DEVELOPMENTAGGREGATIONCELLSSOLUBILIZATIONPURIFICATIONTHROUGHPUTADHESIONCULTURE
제목
Arginine as an Eluent Overcomes the Hindrance of Monoclonal Antibody Quantification by Dextran Sulfate in Protein A Affinity Chromatography
저자
Kim, Bong GyunPark, Hong Woo
DOI
10.1002/btpr.2164
발행일
2015-11
유형
Article
저널명
Biotechnology Progress
31
6
페이지
1536 ~ 1541