Isolation of Hemoglobin from Bovine Erythrocytes by Controlled Hemolysis in the Membrane Bioreactor
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2012
Authors
Stojanović, Radoslava
Ilić, Vesna Lj.

Manojlović, Verica

Bugarski, Diana
Dević, Marija
Bugarski, Branko

Article (Published version)

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In this work, we describe an optimized procedure based on gradual hemolysis for the isolation of hemoglobin derived from bovine slaughterhouse erythrocytes in a membrane bioreactor. The membrane bioreactor system that provided high yields of hemoglobin (mainly oxyhemoglobin derivate) and its separation from the empty erythrocyte membranes (ghosts) was designed at a pilot scale. Ten different concentrations of hypotonic media were assessed from the aspect of the extent of hemolysis, hematocrit values of the erythrocyte suspensions, cell swelling, and membrane deformations induced by decreased salt concentration. Effective gradual osmotic hemolysis with an extent of hemolysis of 88% was performed using 35 mM Na-phosphate/NaCl buffer of pH 7.2-7.4. Under these conditions most of the cell membranes presented the appearance of the normal ghosts under phase contrast microscope. The hemoglobin purity of gt 80% was confirmed by SDS-PAGE. Kinetic studies showed that maximal concentration of he...moglobin was reached after 40 min, but the process cycle at which recovery of 83% was achieved lasted for 90 min. The dynamics of both steps, (1) transport through the membrane of erythrocytes during process of hemolysis and (2) transport through the reactor filters, were evaluated.
Keywords:
Slaughterhouse blood / Bovine erythrocytes / Gradual hemolysis / Membrane bioreactor / Erythrocyte ghostsSource:
Applied Biochemistry and Biotechnology, 2012, 166, 6, 1491-1506Publisher:
- Springer, New York
Funding / projects:
- Novel encapsulation and enzyme technologies for designing of new biocatalysts and biologically active compounds targeting enhancement of food quality, safety and competitiveness (RS-46010)
- EUREKA E!4486
DOI: 10.1007/s12010-012-9543-9
ISSN: 0273-2289
PubMed: 22252906
WoS: 000302770300012
Scopus: 2-s2.0-84858700595
Institution/Community
Tehnološko-metalurški fakultetTY - JOUR AU - Stojanović, Radoslava AU - Ilić, Vesna Lj. AU - Manojlović, Verica AU - Bugarski, Diana AU - Dević, Marija AU - Bugarski, Branko PY - 2012 UR - http://TechnoRep.tmf.bg.ac.rs/handle/123456789/2252 AB - In this work, we describe an optimized procedure based on gradual hemolysis for the isolation of hemoglobin derived from bovine slaughterhouse erythrocytes in a membrane bioreactor. The membrane bioreactor system that provided high yields of hemoglobin (mainly oxyhemoglobin derivate) and its separation from the empty erythrocyte membranes (ghosts) was designed at a pilot scale. Ten different concentrations of hypotonic media were assessed from the aspect of the extent of hemolysis, hematocrit values of the erythrocyte suspensions, cell swelling, and membrane deformations induced by decreased salt concentration. Effective gradual osmotic hemolysis with an extent of hemolysis of 88% was performed using 35 mM Na-phosphate/NaCl buffer of pH 7.2-7.4. Under these conditions most of the cell membranes presented the appearance of the normal ghosts under phase contrast microscope. The hemoglobin purity of gt 80% was confirmed by SDS-PAGE. Kinetic studies showed that maximal concentration of hemoglobin was reached after 40 min, but the process cycle at which recovery of 83% was achieved lasted for 90 min. The dynamics of both steps, (1) transport through the membrane of erythrocytes during process of hemolysis and (2) transport through the reactor filters, were evaluated. PB - Springer, New York T2 - Applied Biochemistry and Biotechnology T1 - Isolation of Hemoglobin from Bovine Erythrocytes by Controlled Hemolysis in the Membrane Bioreactor EP - 1506 IS - 6 SP - 1491 VL - 166 DO - 10.1007/s12010-012-9543-9 ER -
@article{ author = "Stojanović, Radoslava and Ilić, Vesna Lj. and Manojlović, Verica and Bugarski, Diana and Dević, Marija and Bugarski, Branko", year = "2012", abstract = "In this work, we describe an optimized procedure based on gradual hemolysis for the isolation of hemoglobin derived from bovine slaughterhouse erythrocytes in a membrane bioreactor. The membrane bioreactor system that provided high yields of hemoglobin (mainly oxyhemoglobin derivate) and its separation from the empty erythrocyte membranes (ghosts) was designed at a pilot scale. Ten different concentrations of hypotonic media were assessed from the aspect of the extent of hemolysis, hematocrit values of the erythrocyte suspensions, cell swelling, and membrane deformations induced by decreased salt concentration. Effective gradual osmotic hemolysis with an extent of hemolysis of 88% was performed using 35 mM Na-phosphate/NaCl buffer of pH 7.2-7.4. Under these conditions most of the cell membranes presented the appearance of the normal ghosts under phase contrast microscope. The hemoglobin purity of gt 80% was confirmed by SDS-PAGE. Kinetic studies showed that maximal concentration of hemoglobin was reached after 40 min, but the process cycle at which recovery of 83% was achieved lasted for 90 min. The dynamics of both steps, (1) transport through the membrane of erythrocytes during process of hemolysis and (2) transport through the reactor filters, were evaluated.", publisher = "Springer, New York", journal = "Applied Biochemistry and Biotechnology", title = "Isolation of Hemoglobin from Bovine Erythrocytes by Controlled Hemolysis in the Membrane Bioreactor", pages = "1506-1491", number = "6", volume = "166", doi = "10.1007/s12010-012-9543-9" }
Stojanović, R., Ilić, V. Lj., Manojlović, V., Bugarski, D., Dević, M.,& Bugarski, B.. (2012). Isolation of Hemoglobin from Bovine Erythrocytes by Controlled Hemolysis in the Membrane Bioreactor. in Applied Biochemistry and Biotechnology Springer, New York., 166(6), 1491-1506. https://doi.org/10.1007/s12010-012-9543-9
Stojanović R, Ilić VL, Manojlović V, Bugarski D, Dević M, Bugarski B. Isolation of Hemoglobin from Bovine Erythrocytes by Controlled Hemolysis in the Membrane Bioreactor. in Applied Biochemistry and Biotechnology. 2012;166(6):1491-1506. doi:10.1007/s12010-012-9543-9 .
Stojanović, Radoslava, Ilić, Vesna Lj., Manojlović, Verica, Bugarski, Diana, Dević, Marija, Bugarski, Branko, "Isolation of Hemoglobin from Bovine Erythrocytes by Controlled Hemolysis in the Membrane Bioreactor" in Applied Biochemistry and Biotechnology, 166, no. 6 (2012):1491-1506, https://doi.org/10.1007/s12010-012-9543-9 . .