Campus Ring 1 28759 Bremen Germany
Bioprocess intensification and Process proteomics
Downstream processing of natural compounds
Meta-Integration in downstream processing
- Aqueous two-phase extraction (e.g., reactive extraction)
- Membrane adsorbents (e.g., material chemistry, ligand design)
- Expanded bed adsorption (e.g., biomass effects, affinity capture, novel particles)
Continuous processing unit operations for biotechnology Process modeling, simulation, design and scale-up
- Diploma in Biochemistry, University of Buenos Aires (1986)
- Diploma in Industrial Pharmacy and Biochemistry, University of Buenos Aires (1993)
- Dr. in Industrial Microbiology and Biotechnology, University of Buenos Aires (1995)
- Prize "School of Pharmacy and Biochemistry", University of Buenos Aires (1996)
- "Rene Hugo Thalmann" Fellowship, University of Buenos Aires (1996)
- Postdoctoral Fellowship, National Research Council (Argentina) (1997-1999)
- Member of the program "Return of Talents" (CIM/ZAV - BMZ) (2000-2002)
- Postdoctoral Scientist, University of Düsseldorf in the Research Centre Jülich (1997-2000)
- Member of the National Research Council for Science and Technology (Argentina) (since 2000)
- Associate Professor for Biochemical Engineering (Downstream Processing), IUB (since 2003)
Modern biotechnology heavily depends on the availability of efficient processes, which should be able to generate competitive products (or services) in terms of quality and cost. A critical assessment of current bioprocess technology will reveal that fermentation procedures are already in a phase of technological maturity and that many successful approaches to heterologous protein expression are being employed. However, product recovery and purification (also referred as downstream processing) still poses a number of important challenges. For example, downstream operations cost usually represents more than 50-80% of the total processing cost. Thus, it comes as no surprise that optimisation of the later steps is considered a central element in appropriate bioprocess design. Unfortunately, and probably due to the common practice of expressing proteins at laboratory scale without any concern on how the product will be purified at a larger scale, downstream processing strategies are usually neglected until a later phase of process development. This is a major reason for increased processing cost and eventually the reason for product failure in the market (Datar and Rosen, 1990).
The picture described above has fostered the integration between unit operations devoted to product recovery and purification. The mentioned concept materialises through the introduction of newly designed downstream process technologies which replaces a number of more traditional unit operations, like precipitation, centrifugation/filtration, and column chromatography among others. Efforts to perform integrative purification have been made using aqueous two-phase partitioning, membrane adsorbents or fluidised materials for protein capture (Brandt et al., 1988; Kula, 1990; Hjorth, 1997). Moreover, bioprocess integration is a term also used to describe the need to couple properly upstream and downstream operations, something that quite often is left to chance with dramatic consequences (Wielen and Luyben, 1992).
Within the scope of this project, a meta-integration strategy (refer to Fig. 1.) is proposed by full cross-optimisation from gene to product. The prefix meta- is thus "used with the name of a discipline to designate a new but related discipline designed to deal critically with the original one" (Webster Dictionary). As oppose to the standard practice we will start choosing the downstream operation. This will be an adsorption step based on the well known principle of ion exchange (IEX). Chromatography (or the simpler finite batch adsorption) on anion or cation exchangers is a widespread technique for large-scale protein purification, because of economy, robustness and suitability for cleaning in place (CIP) and/or sanitisation in place (SIP). However, ion exchange is usually regarded as a low-to-moderate-resolution purification step. It also requires concentrations of the target species above a certain threshold in order to effectively bind them (Skidmore et al., 1990). On the other hand, we should remember that packed-bed adsorption does not tolerate the presence of particles in the feedstock thus imposing the need for extensive clarification to avoid system clogging and concomitant back-pressure development (Thömmes, 1997). It is clear that a well designed integrative procedure will attempt to simultaneous solid-liquid separation, product selective isolation and de-watering (target concentration). All those potential drawbacks of the chosen core technology (i.e., IEX) are overcome by taking action towards full cross-optimisation of the whole process: a) Use of gene technology to facilitate product purification, i.e. alteration of protein isoelectric point (pI) by addition of charged amino acid residues; b) Selection of the right system with potential to achieve high levels of foreign gene expression, i.e. the baculovirus expression vector system (BEVS); c) Insect cell cultivation in bioreactor(s) for mass protein production, i.e. Stirred Tank vs. Airlift reactors; and d) IEX system, i.e. adsorption on modified hollow fiber membranes vs. adsorption on fluidised and classified matrices.
- Areces L, Biscoglio M, Parry M, Fraile RE, Fernández-Lahore HM and Cascone O. "Purification and characterisation of a milk clotting protease from Mucor bacilliformis" Appl. Biochem. Biotechnol. (1992) 37, 283-291. (*)
- Fernández-Lahore HM, Miranda MV, Fraile ER, Biscoglio M and Cascone O. "Partition behaviour and purification of a Mucor bacilliformis acid protease in aqueous two phase systems" Proc. Biochem. (1995) 30(7), 615-621. (*)
- Miranda MV, Fernández-Lahore HM and Cascone O. "Horseradish peroxidase extraction and purification by aqueous two phase partition" Appl. Biochem. Biotechnol. (1995) 53(2), 147-154.
- Miranda MV, Fernández-Lahore MV and Cascone O. "Influence of the isoelectric point on the thaumatin partition in PEG/Salt aqueous two phase systems" Bioseparations (1996) 6(5), 315-321.
- Fernández-Lahore HM, Gallego-Duaiges MV, Cascone O, Fraile ER. "Solid state production of an acid protease from Mucor bacilliformis" Rev. Arg. Microbiol. (1997) 29, 1-6. (*)
- Grasselli M, Navarro del Cañizo AA, Fernández-Lahore HM, Miranda MV, Camperi SA, and Cascone O. "Qué hacer con el suero de leche" Ciencia Hoy (1997), 8(43), 12-17.
- Fernández-Lahore HM, Fraile ER and Cascone O. "Acid protease recovery from a solid-state fermentation system" J. Biotechnol. (1998) 62, 83-93. (*)
- Miranda MV, Fernandez-Lahore HM, and Cascone O. "Peroxidase extractive purification from plant raw materials in aqueous two-phase systems" Acta Biotechnol. (1998), 18(3), 179-188.
- Fernández-Lahore HM, Auday R, Fraile ER, Biscoglio de Jimenez Bonino M, Pirpignani L, Machalinski C, and Cascone O. "Purification and Characterization of an acid proteinase from mesophilic Mucor spp solid-state cultures" J. Peptide Res. (1999), 53(6), 599-605.
- Fernández-Lahore HM, Kleef R, Kula M.-R, and Thömmes J. "The influence of complex biological feedstock on the fluidisation and bed stability in expanded bed adsorption" Biotech. Bioeng. (1999), 64(4), 484-496. (**)
- Fernández-Lahore HM, Boldt K, Kula M.-R, and Thömmes J. "The influence of cell adsorbent interactions on protein adsorption in expanded beds" J. of Chromatogr. A (2000), 873, 195-208. (**)
- Kijak G, Camperi SA, Stumpo R, Cascone O and Fernández-Lahore HM. "Extractive fractionation of hyperimmune horse sera" Sep. Sci. Technol. (2001), 36(1), 59-79.
- Navarro del Cañizo AA, Fernandez-Lahore HM, Miranda MV, and Cascone O. "Acid protease purification by dye affinity chromatography" Afinidad (2001), 58 (493), 231-233.
- Cabrera R, Stumpo R, López P, Ferraro G, Cascone O, Fernández-Lahore H M. "Protein separation from biological complex mixtures employing continuous liquid-liquid extraction processes" FABICIB (2001), 5, 107-118. (in Spanish)
- Lin D-Q, Fernández-Lahore HM, Kula M-R and Thömmes J. "Minimising Biomass/Adsorbent Interactions in Expanded Bed Adsorption Processes " A Methodological Design Approach" Bioseparation (2001), 10, 7-9. (**)
- Fernández-Lahore H.M., Lin D-Q, Kula M.R., Thömmes J. The use of Ion Selective "Electrodes for evaluating Residence Time Distributions in Expanded Bed Adsorption Systems". Biotech. Progr. (2001), 17(6): 1128-36. (**)
- Magri, L, Cabrera, R, Miranda MV, Fernández-Lahore, HM, Cascone, O. (2003) "Purification of Horseradish peroxidase by counter-current chromatography employing aqueous two-phase systems" Journal of Separation Science, 26/18, 1701-1706.
- Cabrera RB, Saccodossi N, Oliver F, Gelmi L, Fernández-Lahore HM, Leoni J, Stumpo R. (2004) " Chemical extraction and direct adsorptive purification of the recombiant human antigen Ro-52 Journal of Separation Science, 27/7-8, 589-594.
Certificate of Residency Program, Clin. Chem. Microbiol., BUE City Health System (1989)