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Development and characterization of high-performance functionalized membranes for antibody adsorption

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dc.contributorBrazel, Christopher S.
dc.contributorNikles, David E.
dc.contributorClark, Peter E.
dc.contributorBao, Yuping
dc.contributor.advisorRitchie, Stephen M. C.
dc.contributor.authorShethji, Jayraj Kiritbhai
dc.contributor.otherUniversity of Alabama Tuscaloosa
dc.date.accessioned2017-03-01T16:36:06Z
dc.date.available2017-03-01T16:36:06Z
dc.date.issued2012
dc.descriptionElectronic Thesis or Dissertationen_US
dc.description.abstractCapacity and selectivity are the major bottlenecks for the development of affinity membrane adsorbers for protein and antibody purification. The focus of this doctoral research is to develop polyethersulfone (PES) microfiltration (MF) membranes containing multiple highly selective poly(styrene-co-hydroxystyrene) grafts mimicking the key dipeptide Phe-132/Tyr-133 motif of ligand protein A to selectively adsorb immunoglobulin G (IgG) under convective flow conditions. This research work consists of two phases. In phase 1, homopolymer and block copolymer grafts were synthesized and characterized in the membrane pores using monomers styrene, ethoxystyrene, and chloromethylstyrene. 1H NMR characterization showed successful incorporation of the sequential stages of graft chemistry, including: polystyrene, poly(chloromethylstyrene), poly(ethoxystyrene), poly(styrene-b-ethoxystyrene), and poly(styrene-b-chloromethylstyrene). A study of monomer reactivity showed that chloromethylstyrene reacted approximately 1.3 times slower than styrene and ethoxystyrene during formation of homopolymers and block copolymers. The ion-exchange capacity of sulfonated functionalized membranes was 4.9 meq/g with as many as 125 repeat units per chain. In phase 2, PES MF membranes tailored with two different graft chemistries including poly(styrene-co-hydroxystyrene) and glycine functionalized poly((styrene-co-hydroxystyrene)-b-chloromethylstyrene) grafts were developed and tested for selective IgG adsorption. 1H NMR characterization confirmed membrane pore functionalization by poly(styrene-co-hydroxystyrene), chloromethylstyrene block addition, and subsequent glycine functionalization of the chloromethyl block. The dynamic binding capacity (DBC) for IgG was as high as 95 mg/ml, more than 9 times as compared to Sartobind® and Ultrabind® membranes and twice as compared to affinity resin. The DBC was independent of flow rate and there was no significant loss (<5%) in capacity at higher linear velocities (230 cm/h) indicating that the transport of IgG to the adsorptive sites is predominantly by convection. Bind and elute experiments showed that there was no significant loss of DBC over a period of five cycles and the average recovery of antibody was >94%. Competitive sorption using membranes containing negatively charged spacer arms showed that the membrane was ~11 times more selective for IgG than BSA. Additionally, the DBC was 22% higher (115 mg/ml) than without spacer arms indicating that the negatively charged spacer arms moved the grafted chains apart and improved the accessibility of IgG to the binding sites.en_US
dc.format.extent140 p.
dc.format.mediumelectronic
dc.format.mimetypeapplication/pdf
dc.identifier.otheru0015_0000001_0001100
dc.identifier.otherShethji_alatus_0004D_11253
dc.identifier.urihttps://ir.ua.edu/handle/123456789/1580
dc.languageEnglish
dc.language.isoen_US
dc.publisherUniversity of Alabama Libraries
dc.relation.hasversionborn digital
dc.relation.ispartofThe University of Alabama Electronic Theses and Dissertations
dc.relation.ispartofThe University of Alabama Libraries Digital Collections
dc.rightsAll rights reserved by the author unless otherwise indicated.en_US
dc.subjectEngineering
dc.subjectChemical engineering
dc.titleDevelopment and characterization of high-performance functionalized membranes for antibody adsorptionen_US
dc.typethesis
dc.typetext
etdms.degree.departmentUniversity of Alabama. Department of Chemical and Biological Engineering
etdms.degree.disciplineChemical & Biological Engineering
etdms.degree.grantorThe University of Alabama
etdms.degree.leveldoctoral
etdms.degree.namePh.D.

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