title: Influence of different spacer arms on Mimetic Ligand™ A2P and B14 membranes for human IgG purification.
abstract: 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 IgG(1). 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 3.1mg/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 (0.6M sodium hydroxide). Furthermore, foreseeable improvements in binding capacity will promote the implementation of membrane adsorbers in antibody manufacturing.
authors.affiliation: DICMA, Alma Mater Studiorum Università di Bologna, Bologna, Italy. cristiana.boi@unibo.it
authors.forename: Cristiana
authors.initials: C
authors.lastname: Boi
authors.forename: Simone
authors.initials: S
authors.lastname: Dimartino
authors.forename: Stefan
authors.initials: S
authors.lastname: Hofer
authors.forename: Jeannie
authors.initials: J
authors.lastname: Horak
authors.forename: Sharon
authors.initials: S
authors.lastname: Williams
authors.forename: Giulio C
authors.initials: GC
authors.lastname: Sarti
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Lindner
mesh_terms: D000327:Adsorption; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D057930:Click Chemistry; D005019:Ethyl Ethers / Q000737:chemistry; D005029:Ethylenediamines / Q000737:chemistry; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification* / Q000378:metabolism; D008567:Membranes, Artificial; D013438:Sulfhydryl Compounds / Q000737:chemistry; D014230:Triazoles / Q000737:chemistry
chemical_list: C471080:3,6-dioxa-1,8-octanedithiol; D005019:Ethyl Ethers; D005029:Ethylenediamines; D007074:Immunoglobulin G; D008567:Membranes, Artificial; D013438:Sulfhydryl Compounds; D014230:Triazoles; C031234:ethylenediamine