Skip to main content
Log in

High gradient magnetic separation versus expanded bed adsorption: a first principle comparison

  • Published:
Bioseparation

Abstract

A robust new adsorptive separation technique specifically designed for direct product capture from crude bioprocess feedstreams is introduced and compared with the current bench mark technique, expanded bed adsorption. The method employs product adsorption onto sub-micron sized non-porous superparamagnetic supports followed by rapid separation of the ‘loaded’ adsorbents from the feedstock using high gradient magnetic separation technology. For the recovery of Savinase® from a cell-free Bacillus clausii fermentation liquor using bacitracin-linked adsorbents, the integrated magnetic separation system exhibited substantially enhanced productivity over expanded bed adsorption when operated at processing velocities greater than 48 m h−1. Use of the bacitracin-linked magnetic supports for a single cycle of batch adsorption and subsequent capture by high gradient magnetic separation at a processing rate of 12 m h−1 resulted in a 2.2-fold higher productivity relative to expanded bed adsorption, while an increase in adsorbent collection rate to 72 m h−1 raised the productivity to 10.7 times that of expanded bed adsorption. When the number of batch adsorption cycles was then increased to three, significant drops in both magnetic adsorbent consumption (3.6 fold) and filter volume required (1.3 fold) could be achieved at the expense of a reduction in productivity from 10.7 to 4.4 times that of expanded bed adsorption.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Anspach FB, Wirth H-J, Unger KK, Stanton P, Davies JR and Hearn MTW (1989) High performance liquid chromatography with phenylboronic acid, benzamidine, tri-l-alanine and concanavalin A immobilised on 3-isothiocyanatopropyltriethoxysilane-activated nonporous monodisperse silicas. Anal. Biochem. 179: 171–181.

    Google Scholar 

  • Anspach FB, Curbelo D, Hartmann R, Garke G and Deckwer WD (1999) Expanded-bed chromatography in primary protein purification. J. Chromatogr. A 865: 129–144.

    Google Scholar 

  • Barkholt V and Jensen AJ (1989) Amino acid analysis: Determination of cysteine plus half cysteine in protein after hydrochloric acid hydrolysis with a disulphide compound as additive. Anal. Biochem. 177: 318–322.

    Google Scholar 

  • Birrs RR and Parker MR (1981) High intensity magnetic separation. In: Wakeman RJ (ed.) Progress in filtration and separation 2 (pp. 171–303). Elsevier Sci. Publ. Co., Amsterdam.

    Google Scholar 

  • Brummelhuis HGJ (1980) Preparation of the prothrombin complex. In: Curling JM (ed.) Methods of plasma protein fractionation (pp 117–128) Academic Press, London.

    Google Scholar 

  • Chang YK and Chase HA (1996) Development of operating conditions for protein purification using expanded bed techniques: The effect of the degree of bed expansion on adsorption performance. Biotechnol. Bioeng. 49: 512–526.

    Google Scholar 

  • Chase HA (1994) Purification of proteins by adsorption chromatography in expanded beds. Trends Biotechnol. 12: 296–303.

    Google Scholar 

  • Eveleigh JW (1978) Techniques and instrumentation for preparative immunoadsorbent separations. J. Chromatogr. 159: 129–145.

    Google Scholar 

  • Feuser J, Halfar M, Lütkemeyer D, Ameskamp N, Kula MR and Thömmes J (1999) Interaction of mammalian cell culture broth with adsorbents in expanded bed adsorption of monoclonal antibodies. Process Biochemistry 34: 159–165.

    Google Scholar 

  • Franzreb M and Höll WH (2000) Phosphate removal by high gradient magnetic filtration using permanent magnets. IEEE Trans. on Appl. Superconductivity 10, 1: 923–926.

    Google Scholar 

  • Groman EV and Wilchek M (1987) Recent developments in affinity chromatography supports. Trends Biotechnol. 5: 220–224.

    Google Scholar 

  • Hannson M, Stål S, Hjorth R, Uhlen M and Moks T (1994) Single step recovery of a secreted recombinant protein by EBA. Bio/Technology 12: 285–288.

    Google Scholar 

  • Hermanson GT, Malia AK and Smith PK (1992) Immobilised affinity ligand techniques. Academic Press, London.

    Google Scholar 

  • Hjorth R, Kämpe S and Carlsson, M (1995) Analysis of some operating parameters of novel adsorbents for recovery of proteins from expanded bed. Bioseparation 5: 217–224

    Google Scholar 

  • Hjorth R (1997) Expanded bed adsorption in industrial bioprocessing: Recent developments. Trends Biotechnol. 15: 230–235.

    Google Scholar 

  • Karau A, Benken C, Thömmes J and Kula MR (1997) The influence of particle size distribution and operating conditions on the adsorption performance in fluidised beds. Biotechnol. Bioeng. 55 (1): 54–64.

    Google Scholar 

  • Kazumasa Y, Munehiro N, Takao O and Akinori S (1996). Process for purifying recombinant human serum albumin. European patent, EP0699687.

  • Levenspiel O (1999) Chemical Reaction Engineering 3rd edition John Wiley & Sons, New York.

    Google Scholar 

  • Mäkinen KK (1972) Inhibition by bacitracin of some hydrolytic enzymes. Int. J. Protein Research 4: 21–28.

    Google Scholar 

  • Morgan PE (1996) Non-porous pseudo affinity supports for the recovery of antibodies. Ph.D. Thesis, University College of London.

  • Munro PA, Dunnill P and Lilly MD (1977) Nonporous magnetic materials as enzyme supports: studies with immobilised chymotrypsin. Biotechnol. Bioeng. 19: 101–124.

    Google Scholar 

  • O'Brien SM, Thomas ORT and Dunnill P (1996) Non-porous magnetic chelator supports for protein recovery by immobilised metal affinity adsorption. J. Biotechnol. 50: 13–25.

    Google Scholar 

  • O'Brien SM, Sloane RP, Thomas ORT and Dunnill P (1997) Characterisation of non-porous magnetic chelator supports and their use to recover polyhistidine-tailed T4 lysozyme from a crude E. coli extract. J. Biotechnol. 54: 53–67.

    Google Scholar 

  • O'Shannessy K, Scoble J and Scopes RK (1996) A simple and economical procedure for purification of muscle lactate dehydrogenase by batch dye-ligand adsorption. Bioseparation 6: 77–80.

    Google Scholar 

  • Roe SD (1987) Whole broth extraction of enzymes from fermentation broth using commercially available adsorbents. In: Verall MS & Hudson MJ (ed.) Separations for Biotechnology, Ellis Horwood, Chichester.

    Google Scholar 

  • Scoble J, O'Shannessy K and Scopes RK (1996) Purification of recombinant Zymomonas mobilis glucose 6-phosphate dehydrogenase and glucokinase using batch dye-ligand adsorption. Bioseparation 6: 243–246.

    Google Scholar 

  • Scopes RK (1994) Protein Purification-Principle and Practice. 3rd edition Springer, New York.

    Google Scholar 

  • Setchell CH (1985) Magnetic separations in biotechnology-a review. J. Chem. Technol. Biotechnol. 35B: 175–182.

    Google Scholar 

  • Stepanov VM and Rudenskaya GN (1983) Proteinase affinity chromatography on bacitracin Sepharose. J. App. Biochem. 5: 420–428

    Google Scholar 

  • Svoboda J (1987) Magnetic methods for the treatment of minerals. Elsevier Sci. Publ. Co., Amsterdam.

    Google Scholar 

  • Thömmes J, Weiher M, Karau A and Kula MR (1995) Hydrodynamics and performance in fluidised bed adsorption. Biotechnol. Bioeng. 4: 367–374.

    Google Scholar 

  • Thömmes J (1997) Fluidized bed adsorption as a primary recovery step in protein purification. Adv. Biochem. Eng. Biotechnol. 58: 185–230.

    Google Scholar 

  • Watson JHP (1973) Magnetic filtration. J. Appl. Phys. 44: 4209–4213.

    Google Scholar 

  • Whitesides GM, Kazlaukas RJ and Josephson L (1983) Magnetic separations in biotechnology. Trends Biotechnol. 1: 144–148.

    Google Scholar 

  • Zulqarnain K (1999) Scale up of affinity separation based on magnetic support particles. Ph.D. Thesis, University College of London.

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hubbuch, J.J., Matthiesen, D.B., Hobley, T.J. et al. High gradient magnetic separation versus expanded bed adsorption: a first principle comparison. Bioseparation 10, 99–112 (2001). https://doi.org/10.1023/A:1012034923621

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1012034923621

Navigation