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Bioreactor based on suspended particles of immobilized enzyme

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Abstract

A bioreactor for blood detoxification was developed in which oscillation-induced secondary flows suspend particles of immobilized enzyme in a reactor operating at clinically useful flowrates. Torsional oscillation of the reactor about its axis created a pair of counterrotating toroidal vortices which were readily observed in flow-visualization studies. Oscillation frequencies were selected to provide spatially uniform particle dispersion, as assessed visually. As a model system, blood deheparinization by reactors containing heparinase immobilized to agarose particles was investigated. Identical deheparinization profiles were observed in the continuous-flow reactor and in independent batch studies, done in well mixed test tubes of blood, demonstrating that the oscillating reactor design minimizes external mass transfer limitations. Identical heparin neutralization profiles and rates were also observed in the first and the second of consecutive heparin neutralization studies (0–2h and 2–4h, respectively) demonstrating an effective half-life of the immobilized enzyme in the oscillating reactor of at least 4 h. No significant decrease in red or white blood cell count, platelet count, or hematocrit, and clinically acceptable levels of plasma hemoglobin and activated complement were observed with 2 h (20 passes) ofin vitro recirculation of human blood through the reactor. High, stable efficacy, operational stability, and excellent biocompatibility are attributed to secondary flow induced liquid-particle mixing within the oscillating reactor.

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Abbreviations

a :

exponent of least squares fit of heparin neutralization profile:S=S 0 exp(−at)

d p :

particle diameter

E:

enzyme (heparinase) specific activity at timezero (U/cm3 packed gel)

f :

reactor oscillation frequency

Q :

flowrate

Re:

Reynolds number

S :

substrate (heparin) concentration

S 0 :

substrate concentration at time-zero

t 1/2 :

effective half-life of immobilized heparinase

V p :

volume of immobilized enzyme particles

V pl :

total volume of plasma in the system

V p /V pl :

volume fraction of immobilized enzyme particles

v :

flow velocity (cm/s)

μ:

viscosity

ν:

kinematic viscosity

ρ:

density

Θ:

angle through which reactor oscillates (rads)

ω:

angular velocity (rads/s)

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Authors and Affiliations

  1. Division of Health Sciences & Technology, Massachusetts Institute of Technology, Cambridge, MA

    L. E. Freed

  2. Department of Chemical Engineering, Belgrade University, Yugoslavia

    G. V. Vunjak-Novakovic

  3. Department of Biomedical Engineering, Brigham & Women's Hospital, Boston, MA

    P. A. Drinker

  4. Department of Chemical Engineering, Massachusetts Institute of Technology, E25-342, 77 Massachusetts Avenue, 02139, Cambridge, MA

    R. Langer

Authors
  1. L. E. Freed
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  2. G. V. Vunjak-Novakovic
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  3. P. A. Drinker
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  4. R. Langer
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Freed, L.E., Vunjak-Novakovic, G.V., Drinker, P.A. et al. Bioreactor based on suspended particles of immobilized enzyme. Ann Biomed Eng 21, 57–65 (1993). https://doi.org/10.1007/BF02368165

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  • DOI: https://doi.org/10.1007/BF02368165

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