Influence of different spacer arms on Mimetic Ligand™ A2P and B14 membranes for human IgG purification

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• 作者：Cristiana Boi^a ; ^{cristiana.boi@unibo.it"" rel=""nofollow} ; Simone Dimartino^a ; Stefan Hofer^c ; Jeannie Horak^c ; ^{Jeannie.Horak@univie.ac.at"" rel=""nofollow} ; Sharon Williams^b ; Giulio C. Sarti^a ; Wolfgang Lindner^c
• 关键词：Antibody purification ; A2P ; B14 ; Click-Chemistry ; Azide-functionalized membrane ; Affinity membranes
• 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2011
• 出版时间：1 June 2011
• 年：2011
• 卷：879
• 期：19
• 页码：1633-1640
• 全文大小：393 K

文摘

Microporous membranes are an attractive alternative to circumvent the typical drawbacks associated to bead-based chromatography. In particular, the present work intends to evaluate different affinity membranes for antibody capture, to be used as an alternative to Protein A resins. To this aim, two Mimetic Ligands™ A2P and B14, were coupled onto different epoxide and azide group activated membrane supports using different spacer arms and immobilization chemistries. The spacer chemistries investigated were 1,2-diaminoethane (2LP), 3,6-dioxa-1,8-octanedithiol (DES) and [1,2,3] triazole (TRZ). These new mimetic membrane materials were investigated by static and by dynamic binding capacity studies, using pure polyclonal human immunoglobulin G (IgG) solutions as well as a real cell culture supernatant containing monoclonal IgG₁. The best results were obtained by combining the new B14 ligand with a TRZ-spacer and an improved Epoxy 2 membrane support material. The new B14-TRZ-Epoxy 2 membrane adsorbent provided binding capacities of approximately 3.1 mg/mL, besides (i) a good selectivity towards IgG, (ii) high IgG recoveries of above 90 % , (iii) a high Pluronic-F68 tolerance and (iv) no B14-ligand leakage under harsh cleaning-in-place conditions (0.6 M sodium hydroxide). Furthermore, foreseeable improvements in binding capacity will promote the implementation of membrane adsorbers in antibody manufacturing.