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KMID : 1094720120170020309
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Biotechnology and Bioprocess Engineering
2012 Volume.17 No. 2 p.309 ~ p.315
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Bufferless lysis of erythrocytes for isolation of hemoglobin using modified cellulose acetate membranes
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Vanjari Siva Rama Krishna
Deepthi I.
Sandeep K.
Amrutur Bharadwaj
Bhat Navakanta
Srinivasan Sampath
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Abstract
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This paper presents a modified cellulose acetate membrane prepared using a dry casting technique that can be used to perform lysis of erythrocytes and isolation of hemoglobin. Isolation of hemoglobin is thus achieved without the use of lysis buffers. Cellulose acetate (CA) membranes are embedded with ammonium chloride (NH4Cl) and potassium bicarbonate (KHCO3), which act as lysing agents. The presence of embedded salts is confirmed using EDS analysis. The pores in the CA membrane act as filters. The average pore size in these membranes is designed to be 1.5 μM, as characterized by SEM analysis, so that they allow hemoglobin to pass through and block all other cells and unlysed erythrocytes present in blood. When a drop of blood is added to the membrane, the NH4Cl and KHCO3 embedded in the membrane dissolve in plasma and lyse the erythrocytes. The filtered hemoglobin is characterized using UV-Vis Spectroscopy. The results indicate extraction of higher concentration of hemoglobin compared with conventional methods.
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KEYWORD
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erythrocytes, lysis, bufferless, cellulose acetate membranes, filtration, isolation of hemoglobin
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