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  • Kanagarajan, SelvarajuSwedish University of Agricultural Sciences,Sveriges lantbruksuniversitet,Institutionen för växtförädling,Department of Plant Breeding,Sveriges lantbruksuniversitet (SLU),Swedish University of Agricultural Sciences (SLU) (author)

Production of functional human fetal hemoglobin in Nicotiana benthamiana for development of hemoglobin-based oxygen carriers

  • Article/chapterEnglish2021

Publisher, publication year, extent ...

  • Elsevier BV,2021
  • electronicrdacarrier

Numbers

  • LIBRIS-ID:oai:research.chalmers.se:380793aa-d25f-424c-8079-bb75302f25bc
  • https://doi.org/10.1016/j.ijbiomac.2021.06.102DOI
  • https://research.chalmers.se/publication/524887URI
  • https://res.slu.se/id/publ/113252URI
  • https://lup.lub.lu.se/record/061ba15a-115d-4118-93bd-fc1f3af00043URI

Supplementary language notes

  • Language:English
  • Summary in:English

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  • Subject category:art swepub-publicationtype
  • Subject category:ref swepub-contenttype

Notes

  • Hemoglobin-based oxygen carriers have long been pursued to meet clinical needs by using native hemoglobin (Hb) from human or animal blood, or recombinantly produced Hb, but the development has been impeded by safety and toxicity issues. Herewith we report the successful production of human fetal hemoglobin (HbF) in Nicotiana benthamiana through Agrobacterium tumefaciens-mediated transient expression. HbF is a heterotetrameric protein composed of two identical α- and two identical γ-subunits, held together by hydrophobic interactions, hydrogen bonds, and salt bridges. In our study, the α- and γ-subunits of HbF were fused in order to stabilize the α-subunits and facilitate balanced expression of α- and γ-subunits in N. benthamiana. Efficient extraction and purification methods enabled production of the recombinantly fused endotoxin-free HbF (rfHbF) in high quantity and quality. The transiently expressed rfHbF protein was identified by SDS-PAGE, Western blot and liquid chromatography-tandem mass spectrometry analyses. The purified rfHbF possessed structural and functional properties similar to native HbF, which were confirmed by biophysical, biochemical, and in vivo animal studies. The results demonstrate a high potential of plant expression systems in producing Hb products for use as blood substitutes.

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  • Carlsson, Magnus L.R.Swedish University of Agricultural Sciences,Sveriges lantbruksuniversitet,Institutionen för växtförädling,Department of Plant Breeding,Sveriges lantbruksuniversitet (SLU),Swedish University of Agricultural Sciences (SLU) (author)
  • Chakane, SandeepLund University,Lunds universitet,Tillämpad biokemi,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Pure and Applied Biochemistry,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)tbio-spc (author)
  • Kettisen, KarinLund University,Lunds universitet,Tillämpad biokemi,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Pure and Applied Biochemistry,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)tbio-kkn (author)
  • Smeds, EmanuelLund University,Lunds universitet,Antioxidationsmedicin,Forskargrupper vid Lunds universitet,Antioxidation medicine,Lund University Research Groups(Swepub:lu)med-eus (author)
  • Kumar, Ranjeet,1980Chalmers University of Technology(Swepub:cth)ranjeet (author)
  • Ortenlöf, NiklasLund University,Lunds universitet,Infektionsmedicin,Sektion III,Institutionen för kliniska vetenskaper, Lund,Medicinska fakulteten,Infection Medicine (BMC),Section III,Department of Clinical Sciences, Lund,Faculty of Medicine(Swepub:lu)ni6430or (author)
  • Gram, MagnusLund University,Lunds universitet,Pediatrik, Lund,Sektion V,Institutionen för kliniska vetenskaper, Lund,Medicinska fakulteten,Paediatrics (Lund),Section V,Department of Clinical Sciences, Lund,Faculty of Medicine(Swepub:lu)medk-mo0 (author)
  • Åkerström, BoLund University,Lunds universitet,Antioxidationsmedicin,Forskargrupper vid Lunds universitet,Antioxidation medicine,Lund University Research Groups(Swepub:lu)medk-bak (author)
  • Bülow, LeifLund University,Lunds universitet,Tillämpad biokemi,Centrum för tillämpade biovetenskaper,Kemiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Pure and Applied Biochemistry,Center for Applied Life Sciences,Department of Chemistry,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)tbk-lbu (author)
  • Zhu, Li-HuaSwedish University of Agricultural Sciences,Sveriges lantbruksuniversitet,Institutionen för växtförädling,Department of Plant Breeding,Sveriges lantbruksuniversitet (SLU),Swedish University of Agricultural Sciences (SLU)(Swepub:slu)47928 (author)
  • Sveriges lantbruksuniversitetInstitutionen för växtförädling (creator_code:org_t)
  • Sveriges lantbruksuniversitet

Related titles

  • In:International Journal of Biological Macromolecules: Elsevier BV184, s. 955-9660141-81301879-0003

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