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Ing. Michal Gramblička, PhD.
Identification number: 10208
University e-mail: michal.gramblicka [at] stuba.sk
 
Odborný asistent CSc.,PhD. - Department of Chemical and Biochemical Engineering (ICEE FCFT)

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Basic information

Basic information about a final thesis

Type of thesis: Dissertation thesis
Thesis title:Textural and transport properties of adsorbents designed for separation of monoclonal antibodies
Written by (author): Ing. Michal Gramblička, PhD.
Department: Department of Chemical and Biochemical Engineering (ICEE FCFT)
Thesis supervisor: prof. Ing. Milan Polakovič, CSc.
Opponent 1:prof. Ing. Marián Valko, DrSc.
Opponent 2:doc. Ing. Michal Přibyl, PhD.
Opponent 3:Ing. René Faber, PhD.
Final thesis progress:Final thesis was successfully defended.


Additional information

Additional information about the final thesis follows. Click on the language link to display the information in the desired language.

Language of final thesis:English

Slovak        English

Title of the thesis:Textural and transport properties of adsorbents designed for separation of monoclonal antibodies
Summary:The study focuses on experimental determination and model description of textural and transport properties of chromatographic materials designed for separation of large proteins, especially monoclonal antibodies. The investigated porous adsorbents were FractoGel EMD SE HiCap (M) (strong cation exchanger with sulphoethyl functional groups, producer: Merck), SP Sepharose Fast Flow (strong cation exchanger with sulphopropyl functional groups, producer: GE Healthcare), S Ceramic HyperD F (strong cation exchanger with sulphonic functional groups, producer: BioSepra), MabSelect (affinity adsorbent with immobilized protein A, producer: GE Healthcare), rProtein A Sepharose Fast Flow (affinity adsorbent with immobilized protein A, producer: GE Healthcare), Poros 50A High Capacity (affinity adsorbent with immobilized protein A, producer: Applied Biosystems), ProSep --vA High Capacity (affinity adsorbent with immobilized protein A, producer: Millipore), MEP HyperCel (adsorbent with hydrophobic interactions and MEP (mercaptoethylpyridine) as a ligand, producer: BioSepra), Sepabeads FP-DA (weak anion exchanger with diethylamino functional groups, producer: Mitsubishi Chemical) a Sartobind S (grafted cellulose membrane with sulphonic functional groups (a strong cation exchanger), producer: Sartorius). Textural properties were investigated by porosimetric methods of nitrogen adsorption, mercury intrusion, size exclusion of dextran standards, drying and partially also by microscopy. The results of porosimetric measurements served for estimation of characteristic parameters of the porous network, such as the mean pore size and the pore size distribution. Based on the determined pore characteristics, the accessibility of adsorbent for particular proteins was estimated. Proteins used for investigation of the adsorbents were human polyclonal immunoglobulin gamma (Mw 150 000 g/mol, R 5.5 nm), bovine serum albumin (Mw 66 000 g/mol, R 3.3 nm), human serum albumin (Mw 67 000 g/mol, R 3.3 nm), horse skeletal muscle myoglobin (Mw 17 000 g/mol, R 2.1 nm) and fungal fructosyltransferase (Mw 570 000 g/mol, R 10 nm). Transport properties of adsorbents were studied using the method of batch uptake in a stirred vessel and by frontal loading (in case of the chromatographic membrane). The processes were described by suitable mathematic models in order to estimate parameters characterizing the intraparticle transport, such as the effective or pore diffusivity. Some of the main conclusions of the work were following observations and hypotheses: a) adsorbed protein molecules have a dominant role at limitation of the intraparticle diffusive transport in adsorbents with higher binding capacities, b) estimation of the effective diffusivity may be performed with a certain precision without the knowledge of full adsorption isotherm, c) interpretation of the adsorption isotherm parameters depends on consideration of limited protein accessibility to porous structure, d) non-idealities of the breakthrough curve of a protein in an ion-exchanging membrane might be explained by a hypothesis of a parallel adsorption on two types of adsorption sites.
Key words:textural properties, transport properties, adsorbent, diffusion coefficient, hindered diffusion, pore accessibility, protein chromatography, adsorption mechanism

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