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

Kanagarajan, Selvaraju (author)
Swedish University of Agricultural Sciences,Sveriges lantbruksuniversitet,Institutionen för växtförädling,Department of Plant Breeding,Sveriges lantbruksuniversitet (SLU),Swedish University of Agricultural Sciences (SLU)
Carlsson, Magnus L.R. (author)
Swedish University of Agricultural Sciences,Sveriges lantbruksuniversitet,Institutionen för växtförädling,Department of Plant Breeding,Sveriges lantbruksuniversitet (SLU),Swedish University of Agricultural Sciences (SLU)
Chakane, Sandeep (author)
Lund 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
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Kettisen, Karin (author)
Lund 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
Smeds, Emanuel (author)
Lund University,Lunds universitet,Antioxidationsmedicin,Forskargrupper vid Lunds universitet,Antioxidation medicine,Lund University Research Groups
Kumar, Ranjeet, 1980 (author)
Chalmers University of Technology
Ortenlöf, Niklas (author)
Lund 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
Gram, Magnus (author)
Lund 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
Åkerström, Bo (author)
Lund University,Lunds universitet,Antioxidationsmedicin,Forskargrupper vid Lunds universitet,Antioxidation medicine,Lund University Research Groups
Bülow, Leif (author)
Lund 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
Zhu, Li-Hua (author)
Swedish University of Agricultural Sciences,Sveriges lantbruksuniversitet,Institutionen för växtförädling,Department of Plant Breeding,Sveriges lantbruksuniversitet (SLU),Swedish University of Agricultural Sciences (SLU)
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 (creator_code:org_t)
 
Elsevier BV, 2021
2021
English.
In: International Journal of Biological Macromolecules. - : Elsevier BV. - 0141-8130 .- 1879-0003. ; 184, s. 955-966
  • Journal article (peer-reviewed)
Abstract Subject headings
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  • 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.

Subject headings

NATURVETENSKAP  -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
NATURAL SCIENCES  -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
TEKNIK OCH TEKNOLOGIER  -- Industriell bioteknik -- Bioprocessteknik (hsv//swe)
ENGINEERING AND TECHNOLOGY  -- Industrial Biotechnology -- Bioprocess Technology (hsv//eng)
MEDICIN OCH HÄLSOVETENSKAP  -- Medicinsk bioteknologi -- Medicinsk bioteknologi (hsv//swe)
MEDICAL AND HEALTH SCIENCES  -- Medical Biotechnology -- Medical Biotechnology (hsv//eng)
TEKNIK OCH TEKNOLOGIER  -- Industriell bioteknik -- Medicinsk bioteknik (hsv//swe)
ENGINEERING AND TECHNOLOGY  -- Industrial Biotechnology -- Medical Biotechnology (hsv//eng)

Keyword

Oxygen delivery
HBOCs
Plant-made pharmaceuticals
Oxygen therapeutics
Plant molecular farming
Fetal hemoglobin
Heme-binding protein
Fetal hemoglobin
HBOCs
Heme-binding protein
Oxygen delivery
Oxygen therapeutics
Plant molecular farming
Plant-made pharmaceuticals

Publication and Content Type

art (subject category)
ref (subject category)

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