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Purification of recombinant protein A by aqueous two‐phase extraction integrated with affinity precipitation
Author(s) -
Kamihira Masamichi,
Kaul Rajni,
Mattiasson Bo
Publication year - 1992
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260401112
Subject(s) - chromatography , precipitation , aqueous solution , extraction (chemistry) , protein purification , protein precipitation , chemistry , recombinant dna , phase (matter) , aqueous two phase system , solid phase extraction , polymer , affinity chromatography , biochemistry , enzyme , organic chemistry , physics , meteorology , gene
Aqueous two‐phase extraction incorporated affinity precipitation was examined as a technique for protein purification. An enteric coating polymer, Eudragit S100, was employed as a ligand carrier. Eudragit was specifically partitioned to the top phase in the aqueous two‐phase systems. For application of this method to purification of recombinant protein A using human IgG coupled to Eudragit in an aqueous two‐phase system, 80% of protein A added was recovered with 81% purity. The purity was enhanced 26‐fold by thid method. The IgG‐Eudragit could be used repeatedly for the purification process. This seperation method should be applicable to industrial‐scale purification as a new purification procedure combining the advantages and compensating for the disadvantages of the aqueous two‐phase method and affinity precipitation method. © 1992 John Wiley & Sons, Inc.