Convenient method to purify hemoglobin

Hiromi Sakai; Shinji Takeoka; Hiroyuki Nishide; Eishun Tsuchida

doi:10.3109/10731199409117895

Convenient method to purify hemoglobin

Hiromi Sakai, Shinji Takeoka, Hiroyuki Nishide, Eishun Tsuchida

School of Advanced Science and Engineering

Research output: Contribution to journal › Article › peer-review

8 Citations (Scopus)

Abstract

A convenient method to purify Hb solution from outdated RBC has been established for the starting material of Hb-based blood substitutes. To prevent MetHb formation during the procedure, Hb in RBC was carbonylated in advance. Then RBC was mixed with organic solvent for hemolysis and centrifuged for removal of stroma. The resulting SFHb solution was heated at 60C and generated precipitates were removed out by centrifugation. The purity of Hb (25 g/dl) was confirmed by SDS-PAGE. IEF and oxygen binding property of the Hb solution also guaranteed its purity and no denaturation of Hb. This method is applicable to large scale production of the purified Hb for the starting material of Hb-based blood substitutes.

Original language	English
Pages (from-to)	651-656
Number of pages	6
Journal	Artificial Cells, Blood Substitutes, and Biotechnology
Volume	22
Issue number	3
DOIs	https://doi.org/10.3109/10731199409117895
Publication status	Published - 1994 Jan

ASJC Scopus subject areas

Biotechnology
Biomedical Engineering

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author = "Hiromi Sakai and Shinji Takeoka and Hiroyuki Nishide and Eishun Tsuchida",

year = "1994",

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AU - Sakai, Hiromi

AU - Takeoka, Shinji

AU - Nishide, Hiroyuki

AU - Tsuchida, Eishun

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N2 - A convenient method to purify Hb solution from outdated RBC has been established for the starting material of Hb-based blood substitutes. To prevent MetHb formation during the procedure, Hb in RBC was carbonylated in advance. Then RBC was mixed with organic solvent for hemolysis and centrifuged for removal of stroma. The resulting SFHb solution was heated at 60C and generated precipitates were removed out by centrifugation. The purity of Hb (25 g/dl) was confirmed by SDS-PAGE. IEF and oxygen binding property of the Hb solution also guaranteed its purity and no denaturation of Hb. This method is applicable to large scale production of the purified Hb for the starting material of Hb-based blood substitutes.

AB - A convenient method to purify Hb solution from outdated RBC has been established for the starting material of Hb-based blood substitutes. To prevent MetHb formation during the procedure, Hb in RBC was carbonylated in advance. Then RBC was mixed with organic solvent for hemolysis and centrifuged for removal of stroma. The resulting SFHb solution was heated at 60C and generated precipitates were removed out by centrifugation. The purity of Hb (25 g/dl) was confirmed by SDS-PAGE. IEF and oxygen binding property of the Hb solution also guaranteed its purity and no denaturation of Hb. This method is applicable to large scale production of the purified Hb for the starting material of Hb-based blood substitutes.

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Convenient method to purify hemoglobin

Abstract

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