The amazing ability of continuous chromatography to adapt to a moving environment.
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1 The amazing ability of continuous chromatography to adapt to a moving environment. Roger-Marc Nicoud, Founder of Novasep Barcelona, October 2013
2 Introduction My thesis: the environment pressure pushes Continuous Chromatography to evolve like animals evolved according to Darwin s law. and for that, I will use shortcuts, approximations and manipulations
3 Improving what? Counter-current can help Continuous can help Comment System size Productivity kg/kg/day normally presented as the main objective function Eluent consumption Minimizing solvent consumption (prior to recycling) is a must in all large scale applications. Yield Can be a major cost contributor Operations Control / Process Stability / Cleaning can be critical costs contributors. Mitigation large LP systems can be cheaper than smaller HP systems Biopharma industry still a bit less concerned Ancillary equipment is even more important than chromatography itself. Life is often a matter of compromize
4 Continuous is NOT synonymous with counter-current A continuous but NOT counter-current animal
5 Be careful with comparisons Productivity kg/kg/d Optimum particle size, eluent composition are different for Batch and continuous/cc processes. 2 0 Batch My clever process
6 S.M.B.: the key evolution.. Evolution lead to the first continuous AND counter-current animal solid liquid Maximize productivity IV Feed III II Eluent I
7 The first evidence of continuous chromatography
8 Environment pressure : Paraxylene Scale (500,000 Mt/Y/Unit in 2010) Chromatographic conditions: Selectivity about 2 Particle size about 1000 microns (inducing poor HETP.) Temperature about 180 C (inducing low viscosity) Technical constraints Target purity: > 99.8 % individual bed length : limited to 1 m due to mechanical constraints significant bed length of 24 m (no pb because of low viscosity) 24 columns to reach total length
9 Environment pressure : Fructose Scale (50,000 Mt/Y/Unit in 2010): from Glucose-Fructose mixtures Chromatographic conditions: Selectivity about 1.5 Particle size about microns (inducing poor HETP) Temperature: 60 C (thus viscosity about 10 cp ) Technical constraints Target purity: about 90 % Minimize energy consumption for evaporating Water total bed length of about 8-12 meters
10 Improving the classical SMB Add some degree of freedom by: Connecting columns in different ways Selecting different switching modes Varying I/O lines composition and flows New processes are normally: quasi-continuous instead of being truly continuous taking advantage of «counter-current» contact
11 Fructose: further improvement A new animal Two sub periods «Cousin animals»: SSMB Novasep ISMB Mitsubishi Commercial solutions implemented: 4 or 6 columns Eluent consumption decreased by about % compared to SMB
12 Environment pressure : optical isomers Scale (1-100 Mt/Y/Unit in 2010) Chromatographic conditions: Selectivity: typically between 1.2 and 2 Little influence of particle size on packing cost HETP depending on particle size Technical constraints Target purity: % Minimize system size / CSP inventory Use small particle size Total bed length about 0.5 m. To be distributed between Ncol At large scale L/D < 0.1..
13 The first SMB animal for separating optical isomers (1992): Influence of petrochemical design is very visible. and the modern version.
14 Optical isomers: further improvement VariCol VariCol: Truly continuous Same hardware as SMB Typically saves one column out of 5-6.
15 Key successes For main successes, continuous chromatography is coupled with crystallization, isomerization and multiple effect evaporators
16 Very large scale does not mean «second-class» performance
17 In Line Dilution is key
18 Environment pressure : need for multicomponent (bio)separations. The generic purification problem : An animal with no progeny? Prefer two SMB in series : Paclitaxel, Cyclosporine, time EPA W P S reduces to a binary separation if one wants the green or change or the concept red product. SMB is perfectly adapted to these situations (ex.: desalting, xylenes, capture ), VIII VII VI V IV III II I Technically one can design a continuous three pure fractions TMB. Imposes to work with two suboptimal systems?
19 Improving multicomponent separations VI MCSGP (Multicolumn Countercurrent Solvent Gradient Purification) from ETH. W P P+W Feed V IV W Important features of the animal: Opening liquid loop Short-circuit Gradient S P+S III P A truly continuous gradient chromatographic process with 6-columns II I S
20 3-column MCSGP Two sub periods VI VI P+W V W P+W V P+S IV Cousin animal from Novasep : GSSR IV Gradient (with) Steady-State Recycling P P+S III III Can be done with three columns II I S II I Not strictly continuous but counter current
21 Environment pressure : need for capturing (biomolecules) Selectivity is such that a solvent change is mandatory t Loading Washing Elution W Non retained P S Very strongly retained t cycle Regeneration Equilibration
22 Bioseparations: the BioSc concept Two sub periods W W Feed Wash Elute Regenerate Equilibrate Cousin W animals from : Chromacon (Capture SMB) GE (3C PCC) Wash Target P S Wash * Elute Regenerate Waste Equilibrate W P S Waste Target Use sequential feeding to reduce the number columns (as low as two).
23 Lysine production main steps FERMENTATION MEMBRANE FILTRATION BIOMASS ION EXCHANGE PURIFICATION ACIDIFICATION About 50,000 MT/year/unit EVAPORATION CRISTALLISATION CRUDE L LYSINE HCl
24 Lysine purification: switch from batch to continous (a decade ago) First step: Strong Cation Exchange Feed ph 3-4 so that Lys + (maximizes loading and separation) Then ph lowered to 1-2 so that Lys ++ (maximizes capture of small concentrations) Wash Elution by ammonia at ph 9 (Lys 0 ) Ammonia recycled through stripping 97% Lys with microbial byproducts and mineral cations Second step: Weak Cation Exchange Feed at ph 9 (Lys 0 goes through) Regeneration at ph with diluted sulphuric acid Wash Gain continuous (about 10-column systems) vs Batch: resin inventory and effluent divided by about 3
25 Biopharma Many proof of concept published: mabs pegylated proteins peptides fatty acids.. Still looking for a first Biopharma industrial application of continuous chromatography?
26 Evolution. Xylene 24 Continuous character 100 % Number of columns 12 Sugars I-SMB S-SMB 6 VariCol MCSGP 4 MCSGP GSSR BioSc 3C PCC 2 50 % ?
27 Thank you!
28 Process integration may lead to non intuitive processes Intuition Glu 95 % Isomerization Glu-Fru S M B Glu 90 % HFS 55 % Optimized Glu 95 % Isomerization Glu-Fru S M B Glu 90 % Fru 90 % HFS 55 %
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