Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

Metal chelate affinity precipitation: purification of (His)6-tagged lactate dehydrogenase using poly(vinylimidazole-co-N-isopropylacrylamide)copolymers

Kumar, Ashok LU ; Khalil, Ashraf LU ; Galaev, Igor LU and Mattiasson, Bo LU (2003) In Enzyme and Microbial Technology 33(1). p.113-117
Abstract
Affinity precipitation is a highly selective bioseparation method, which has been successfully used in the purification of various enzymes/proteins. The separation of (His)6-tagged lactate dehydrogenase ((His)6-LDH) was carried through affinity precipitation technique using thermoresponsive copolymer poly(vinylimidazole-co-N-isopropyl-acrylamide) (poly(VI-NIPAM)). Nearly quantitative recovery of (His)6-LDH has been achieved using Cu(II)- and Ni(II)-loaded poly(VI-NIPAM) with Cu(II)-poly(VI-NIPAM) being more efficient in enzyme precipitation than Ni(II)-poly(VI-NIPAM). Optimal precipitation of (His)6-LDH was achieved at pH 6 and pH 6.5–7.0 with Cu(II)-poly(VI-NIPAM) and Ni(II)-poly(VI-NIPAM), respectively. Furthermore, the enzyme was... (More)
Affinity precipitation is a highly selective bioseparation method, which has been successfully used in the purification of various enzymes/proteins. The separation of (His)6-tagged lactate dehydrogenase ((His)6-LDH) was carried through affinity precipitation technique using thermoresponsive copolymer poly(vinylimidazole-co-N-isopropyl-acrylamide) (poly(VI-NIPAM)). Nearly quantitative recovery of (His)6-LDH has been achieved using Cu(II)- and Ni(II)-loaded poly(VI-NIPAM) with Cu(II)-poly(VI-NIPAM) being more efficient in enzyme precipitation than Ni(II)-poly(VI-NIPAM). Optimal precipitation of (His)6-LDH was achieved at pH 6 and pH 6.5–7.0 with Cu(II)-poly(VI-NIPAM) and Ni(II)-poly(VI-NIPAM), respectively. Furthermore, the enzyme was purified by affinity precipitation upto 3.5- and 3.1-fold with recoveries of 95 and 82% using Cu(II)- and Ni(II)-loaded copolymers, respectively. (Less)
Please use this url to cite or link to this publication:
author
; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Affinity precipitation, Lactate dehydrogenase, Thermoresponsive copolymers, Metal affinity
in
Enzyme and Microbial Technology
volume
33
issue
1
pages
113 - 117
publisher
Elsevier
external identifiers
  • wos:000183967800016
  • scopus:0038072717
ISSN
0141-0229
DOI
10.1016/S0141-0229(03)00092-9
language
English
LU publication?
yes
id
ea5de19d-0e9e-4ed9-9c49-d217f2cfee62 (old id 129156)
date added to LUP
2016-04-01 12:34:30
date last changed
2025-10-14 10:36:38
@article{ea5de19d-0e9e-4ed9-9c49-d217f2cfee62,
  abstract     = {{Affinity precipitation is a highly selective bioseparation method, which has been successfully used in the purification of various enzymes/proteins. The separation of (His)6-tagged lactate dehydrogenase ((His)6-LDH) was carried through affinity precipitation technique using thermoresponsive copolymer poly(vinylimidazole-co-N-isopropyl-acrylamide) (poly(VI-NIPAM)). Nearly quantitative recovery of (His)6-LDH has been achieved using Cu(II)- and Ni(II)-loaded poly(VI-NIPAM) with Cu(II)-poly(VI-NIPAM) being more efficient in enzyme precipitation than Ni(II)-poly(VI-NIPAM). Optimal precipitation of (His)6-LDH was achieved at pH 6 and pH 6.5–7.0 with Cu(II)-poly(VI-NIPAM) and Ni(II)-poly(VI-NIPAM), respectively. Furthermore, the enzyme was purified by affinity precipitation upto 3.5- and 3.1-fold with recoveries of 95 and 82% using Cu(II)- and Ni(II)-loaded copolymers, respectively.}},
  author       = {{Kumar, Ashok and Khalil, Ashraf and Galaev, Igor and Mattiasson, Bo}},
  issn         = {{0141-0229}},
  keywords     = {{Affinity precipitation; Lactate dehydrogenase; Thermoresponsive copolymers; Metal affinity}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{113--117}},
  publisher    = {{Elsevier}},
  series       = {{Enzyme and Microbial Technology}},
  title        = {{Metal chelate affinity precipitation: purification of (His)6-tagged lactate dehydrogenase using poly(vinylimidazole-co-N-isopropylacrylamide)copolymers}},
  url          = {{http://dx.doi.org/10.1016/S0141-0229(03)00092-9}},
  doi          = {{10.1016/S0141-0229(03)00092-9}},
  volume       = {{33}},
  year         = {{2003}},
}