Development of Time-course Oxygen Binding Analysis for Hemoglobin-based Oxygen Carriers

Abstract

Developing blood substitutes is in urgent demand for chronic blood shortage all over the world. In this connection, the oxygen binding behavior of hemoglobin-based oxygen carriers (HBOCs) is one of the most important characteristics. However, present methods available for estimating oxygen binding behavior have need of expensive apparatus, and also are not suitable for high-throughput and the time-course analysis. To overcome these problems, we proposed a simple analysis method for the time-course oxygen binding behavior of HBOCs, which employs a general UV-Vis microplate reader and a common reagent, sodium dithionite, as a reductant for HBOCs and an oxygen scavenger. Our method enabled time-course oxygen binding behavior analysis of HBOCs in a simple manner, and obtained data corresponding with those by the conventional method. Thus, our developed method will accelerate the development of HBOCs due to easy oxygen binding analysis.

Biosynthesis of High‐Active Hemoproteins by the Efficient Heme‐SupplyPichia PastorisChassis

“…S at.O2 stands for oxygen saturation degree. [ 55 ] f) HPLC analysis of the catalytic efficiency of cells expressing BM3 mut in converting phenol to hydroquinone. Data presented as mean values ± SD from three independent biological replicates ( n = 3).…”

“…The oxygen binding measurement of V‐Hb was reported as the time course of S at.O2 (oxygen saturation degree). [ 55 ] $$$$\begin{eqnarray} {S}_{{\mathrm{at}}{\mathrm{.O2}}} &=& \frac{{{K}_{{\mathrm{Hb}}} - {K}_{\mathrm{x}}}}{{{K}_{{\mathrm{Hb}}} - {K}_{{\mathrm{HbO2}}}}},\nonumber\\ {K}_{{\mathrm{Hb}}} &=& \frac{{{A}_{{\mathrm{556}}}}}{{{A}_{{\mathrm{523t = 0}}}}},{K}_{{\mathrm{HbO2}}} = \frac{{{A}_{556}}}{{{A}_{523{\mathrm{t = }}\infty }}},{K}_x = \frac{{{A}_{{\mathrm{556}}}}}{{{A}_{{\mathrm{523t = t}}}}} \end{eqnarray}$$$$…”

“…The oxygen binding measurement of V‐Hb was reported as the time course of S at.O2 (oxygen saturation degree). [ 55 ] …”

“…e) The time-course oxygen binding behavior of V-Hb in the presence of sodium dithionite (reductant and oxygen scavenger). S at.O2 stands for oxygen saturation degree [55]. f) HPLC analysis of the catalytic efficiency of cells expressing BM3 mut in converting phenol to hydroquinone.…”