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Activation of Supports Containing Hydroxyl Groups Using bis(4-Nitrophenyl) carbonate

 

作者: Dyer Narinesingh,   T. Ngo,  

 

期刊: Analytical Letters  (Taylor Available online 1996)
卷期: Volume 29, issue 4  

页码: 547-564

 

ISSN:0003-2719

 

年代: 1996

 

DOI:10.1080/00032719608000420

 

出版商: Taylor & Francis Group

 

关键词: bis(4-nitrophenyl) carbonate activated supports;IgG purification;affinity chromatography

 

数据来源: Taylor

 

摘要:

The hydroxyl groups carrying supports, Sepharose Cl-4B and Fractogel HW75F, were facilely activated under anhydrous conditions using bis(4-nitrophenyl) carbonate (BPNPC), a stable and readily available compound. The activation time at room temperature can be as short as 0.5 hour. The activated supports can be stored at 4°C for at least one month without any loss of coupling capacity. Amines can be coupled to the activated support at pH 7 to 10. Coupling of Protein A to BPNPC activated supports yielded immobilized Protein A affinity supports having a high IgG binding capacity. For example, the Protein A-Sepharose and Protein A-Fractogel prepared by using the BPNPC activation method were able to bind, respectively 13 - 14 mg and 8 - 9 mg human IgG with phosphate buffer saline as the loading buffer. In contrast the use of a high yield binding buffer increases the binding capacity of the two affinity supports to 25 - 28 mg and 22 mg respectively.

 

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