Purification of Concentrated Hemoglobin Using Organic Solvent and Heat Treatment

H. Sakai, Shinji Takeoka, H. Yokohama, Y. Seino, Hiroyuki Nishide, E. Tsuchida

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

A simple method for obtaining a purified and concentrated hemoglobin (Hb) solution (25 g/100 ml) from human red blood cells has been established. To prevent MetHb formation during the purification procedure, Hb in red blood cells was carbonylated in advance, and then washed red blood cells were mixed with organic solvents such as diethyl ether or dichloromethane for hemolysis and removal of stroma. The Hb solution was isolated by centrifugation (1 900g) with the high removal efficiency of phospholipid (>99.8%). After the solution was heated (60°C, 1 h), the precipitates were removed by centrifugation. The purity of Hb was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Isoelectric focusing and oxygen-binding properties of the obtained Hb solution demonstrated its purity and showed no denaturation of globin. This purification procedure is applicable to large-scale production of the purified Hb.

Original languageEnglish
Pages (from-to)563-569
Number of pages7
JournalProtein Expression and Purification
Volume4
Issue number6
DOIs
Publication statusPublished - 1993 Dec

Fingerprint

Organic solvents
Purification
Hemoglobins
Hot Temperature
Heat treatment
Blood
Centrifugation
Erythrocytes
Cells
Denaturation
Globins
Methylene Chloride
Isoelectric Focusing
Hemolysis
Electrophoresis
Sodium Dodecyl Sulfate
Ether
Precipitates
Polyacrylamide Gel Electrophoresis
Phospholipids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Purification of Concentrated Hemoglobin Using Organic Solvent and Heat Treatment. / Sakai, H.; Takeoka, Shinji; Yokohama, H.; Seino, Y.; Nishide, Hiroyuki; Tsuchida, E.

In: Protein Expression and Purification, Vol. 4, No. 6, 12.1993, p. 563-569.

Research output: Contribution to journalArticle

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