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Applied Biochemistry and Biotechnology

, Volume 68, Issue 1–2, pp 121–133 | Cite as

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
  • I. Yu. Galaev
  • A. Kumar
  • R. Agarwal
  • M. N. Gupta
  • B. MattiassonEmail author
Original Articles

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.

Index Entries

Immobilized metal affinity precipitation Kunitz soybean inhibitor poly(N-isopropylacrylamide) 

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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Copyright information

© Humana Press Inc 1997

Authors and Affiliations

  • I. Yu. Galaev
    • 1
  • A. Kumar
    • 2
  • R. Agarwal
    • 2
  • M. N. Gupta
    • 2
  • B. Mattiasson
    • 1
    Email author
  1. 1.Department of Biotechnology, Center for Chemistry and Chemical EngineeringLund UniversityLundSweden
  2. 2.Chemistry DepartmentIndian Institute of TechnologyNew DelhiIndia

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