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Imidazole—a new ligand for metal affinity precipitation

Precipitation of Kunitz soybean trypsin inhibitor using cu(II)-loaded copolymers of 1-vinylimidazole withN-vinylcaprolactam orN-isopropylacrylamide

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Abstract

Kunitz soybean trypsin inhibitor (STI) was specifically coprecipitated during precipitation of Cu(II)-loaded copolymers induced by increase in temperature and ionic strength. The copolymers used consisted of 1-vinylimidazole andN-vinylcaprolactam orN- isopropylacrylamide. The elution of STI was achieved by solubilization of the STI-Cu(II)-polymer complex in the presence of an excess of the competing ligand, imidazole, and a subsequent precipitation of the polymer with STI remaining free in solution in a purified form as judged by Sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). To the best of our knowledge this is the first reported successful metal affinity precipitation of protein in a heterobifunctional format.

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Abbreviations

IDA:

iminodiacetic acid

IMAC:

immobilized metal affinity chromatography

VI:

1-vinylimidazole

poly-VI:

poly(l-vinylimidazole)

VCL:

N-vinyl caprolactam

poly-VCL:

poly(N-vinyl caprolactam)

NIPAM:

N-sopropylacrylamide

poly-NIPAM:

poly(N-isopropylacrylamide)

poly-VI-VCL:

copolymer of VI with VCL

poly-VI-NIPAM:

copolymer of VI with NIPAM

BSA:

bovine serum albumin

IgG:

immunoglobulin gamma

STI:

Kunitz soybean trypsin inhibitor

AIBN:

2,2t’-azobisisobutyronitrile

TEMED:

tetraethylene methylenediamine

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Authors and Affiliations

  1. Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, S-22100, Lund, Sweden

    I. Yu. Galaev & B. Mattiasson

  2. Chemistry Department, Indian Institute of Technology, Hauz Khas, 110016, New Delhi, India

    A. Kumar, R. Agarwal & M. N. Gupta

Authors
  1. I. Yu. Galaev
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  2. A. Kumar
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  3. R. Agarwal
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  4. M. N. Gupta
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  5. B. Mattiasson
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Corresponding author

Correspondence to B. Mattiasson.

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Galaev, I.Y., Kumar, A., Agarwal, R. et al. Imidazole—a new ligand for metal affinity precipitation. Appl Biochem Biotechnol 68, 121–133 (1997). https://doi.org/10.1007/BF02785985

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  • DOI: https://doi.org/10.1007/BF02785985

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