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Chromatographic separation of hemoglobin variants using robust molecularly imprinted polymers

Zhang, Ka LU ; Zhou, Tongchang LU ; Kettisen, Karin LU ; Ye, Lei LU and Buelow, Leif LU (2019) In Talanta 199. p.27-31
Abstract
Devising a robust, efficient and cost effective hemoglobin (Hb) purification strategy is one of the key challenges in the development of Hb-based blood substitutes. The aim of this study was to use molecularly imprinted polymers (MIPs) as a novel and efficient chromatographic resin to selectively recognize and purify different Hb variants. The results showed that the Hb-MIP material developed here could selectively recognize and purify various Hb directly from either crude E. coli extracts or human body fluids, such as blood plasma and cerebrospinal fluid (CSF), in one-step. The dynamic binding capacity at 10% breakthrough was around 7.4 mg mL−1resin for adult Hb (HbA) and fetal Hb (HbF). This chromatographic material also allowed... (More)
Devising a robust, efficient and cost effective hemoglobin (Hb) purification strategy is one of the key challenges in the development of Hb-based blood substitutes. The aim of this study was to use molecularly imprinted polymers (MIPs) as a novel and efficient chromatographic resin to selectively recognize and purify different Hb variants. The results showed that the Hb-MIP material developed here could selectively recognize and purify various Hb directly from either crude E. coli extracts or human body fluids, such as blood plasma and cerebrospinal fluid (CSF), in one-step. The dynamic binding capacity at 10% breakthrough was around 7.4 mg mL−1resin for adult Hb (HbA) and fetal Hb (HbF). This chromatographic material also allowed identification of changes related to amino acid substitutions on the Hb protein surface. For instance, when an additional lysine residue was introduced, the HbA αY42K mutant eluted later in an Hb-MIP column than wildtype HbA. Additional negative charges on the protein surface, such as aspartate, mitigated the interaction between the protein and imprinted polymers, and therefore an αA19D-αA12D HbF mutant eluted earlier, at −2.7 column volumes compared to wildtype HbF. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Talanta
volume
199
pages
27 - 31
publisher
Elsevier
external identifiers
  • scopus:85061533637
  • pmid:30952256
ISSN
1873-3573
DOI
10.1016/j.talanta.2019.01.125
language
English
LU publication?
yes
id
a75b87fe-85a9-46dd-bdc8-95d966df6c27
date added to LUP
2019-02-18 13:59:24
date last changed
2020-02-19 05:20:47
@article{a75b87fe-85a9-46dd-bdc8-95d966df6c27,
  abstract     = {Devising a robust, efficient and cost effective hemoglobin (Hb) purification strategy is one of the key challenges in the development of Hb-based blood substitutes. The aim of this study was to use molecularly imprinted polymers (MIPs) as a novel and efficient chromatographic resin to selectively recognize and purify different Hb variants. The results showed that the Hb-MIP material developed here could selectively recognize and purify various Hb directly from either crude E. coli extracts or human body fluids, such as blood plasma and cerebrospinal fluid (CSF), in one-step. The dynamic binding capacity at 10% breakthrough was around 7.4 mg mL−1resin for adult Hb (HbA) and fetal Hb (HbF). This chromatographic material also allowed identification of changes related to amino acid substitutions on the Hb protein surface. For instance, when an additional lysine residue was introduced, the HbA αY42K mutant eluted later in an Hb-MIP column than wildtype HbA. Additional negative charges on the protein surface, such as aspartate, mitigated the interaction between the protein and imprinted polymers, and therefore an αA19D-αA12D HbF mutant eluted earlier, at −2.7 column volumes compared to wildtype HbF.},
  author       = {Zhang, Ka and Zhou, Tongchang and Kettisen, Karin and Ye, Lei and Buelow, Leif},
  issn         = {1873-3573},
  language     = {eng},
  pages        = {27--31},
  publisher    = {Elsevier},
  series       = {Talanta},
  title        = {Chromatographic separation of hemoglobin variants using robust molecularly imprinted polymers},
  url          = {http://dx.doi.org/10.1016/j.talanta.2019.01.125},
  doi          = {10.1016/j.talanta.2019.01.125},
  volume       = {199},
  year         = {2019},
}