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Protein purification by counteracting chromatographic electrophoresis — The focusing window

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Abstract

Counteracting Chromatographic Electrophoresis (CACE) is a novel purification technique that focuses the target protein into a thin band at the interface separating two gel media which differ sharply in internal porosity. This article validates a criterion that predicts protein focusing by CACE, using the colored proteins-ferritin, hemoglobin and myoglobin. Fair comparison is shown between the theory and the experiment. Data from the past literature which were reported to exhibit nonconformity have also been shown to agree fairly well with the present model. Variations in protein focusing conditions can be partly explained by protein-ion binding.

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Abbreviations

c i g/cm3 :

concentration of protein in the interstitial fluid

c 0 g/cm3 :

a characteristic concentration in the CACE system

L cm:

length of the column

t min:

time coordinate

u (cm/min)/(V/cm):

free electrophoretic mobility

u g (cm/min)/(V/cm):

electrophoretic mobility in the gel

v cm/min:

convective velocity

x cm:

axial coordinate

X :

axial coordinate

α :

ratio of accessible volume to inaccessible volume in the column with respect to the target protein (dimensionless)

β :

internal porosity of the gel beads (dimensionless)

ɛ :

external porosity of the packed bed (dimensionless)

φ V/cm:

potential gradient

μ :

ratio of electrophoretic velocity to convective velocity of the target protein (dimensionless)

ψ :

ratio of electrophoretic mobilities (dimensionless)

τ :

time (dimensionless)

ex :

refers to the physical property in the excluding gel region

in :

refers to the physical property in the including gel region

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Authors and Affiliations

  1. Centre for Research & Development Southern Petrochemical Industries Corporation, Tuticorin-628005 Tamilnadu, India

    C. B. Chidambara Raj

  2. Department of Agricultural and Biological Engineering, Cornell University, 14853, Ithaca, New York, USA

    C. B. Chidambara Raj & J. B. Hunter

Authors
  1. C. B. Chidambara Raj
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  2. J. B. Hunter
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Chidambara Raj, C.B., Hunter, J.B. Protein purification by counteracting chromatographic electrophoresis — The focusing window. Bioprocess Engineering 8, 121–128 (1992). https://doi.org/10.1007/BF01254227

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  • DOI: https://doi.org/10.1007/BF01254227

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