Improved oxygen storage capacity of haemoglobin submicron particles by one-pot formulation

Abstract

The coprecipitation-cross-linking-dissolution (CCD) technique for protein submicron particle fabrication was improved by omitting one preparation step using the macromolecular cross-linker, periodate-oxidized dextran (Odex, M.W. of 40 and 70 kDa). The coprecipitation and cross-linking of haemoglobin (Hb) were combined in one single step since the cross-linker is incorporated into the inorganic template, MnCO 3 , together with the protein. After removal of the MnCO 3 templates by EDTA, the amount of entrapped Hb was 60 to 70% of the initial amount. This technique provides deformable Hb submicron particles (HbMP) with narrow size distribution between 800 and 1000 nm, uniform morphology and negative zeta-potential. More than 40% of Hb in the particles was able to carry oxygen over a storage period of 90 days. The results suggest that our new protein submicron particle fabrication technique minimizes the fabrication time and is very efficient and cost-effective.

Künstliche Sauerstofftransporter können mehr als Sauerstoff liefern

“…B. mittels Glutaraldehyds. Die beiden Schritte der Präzipitation und Vernetzung lassen sich bei Verwendung von oxidiertem Dextran zu einem Schritt zusammenfassen [89,90]. Die schwammartigen HbMP sind das Endergebnis nach der Auflösung des anorganischen Carbonat-Templates.…”

Dextran Aldehyde in Biocatalysis: More Than a Mere Immobilization System

“…Finally, it is worth mentioning the use of dextran-hemoglobin conjugates as blood substitutes for patients suffering from asthma or other hemoglobin-compromised conditions. These dextran derivatives not only increase the therapeutic efficiency by enhancing the binding and release of oxygen [111][112][113][114], but also prevent hemoglobin excretion (avoiding any kidney damage [115,116]), reduce the non-renal clearance of hemoglobin (therefore minimizing risks of tissue edema [117]) and reduce the auto-oxidation described for other hemoglobin-based oxygen carriers (reducing tissue and cellular toxicity [118]).…”

Surface Modification of Hemoglobin-Based Oxygen Carriers Reduces Recognition by Haptoglobin, Immunoglobulin, and Hemoglobin Antibodies

“…The total hemoglobin concentration of the surface-modified HbMPs was determined by the modified alkaline hematin-D method (AHD method) [26,27]. Briefly, 2% (v/v) particle suspensions were mixed with pronase to obtain a final concentration of 0.5 mg/mL pronase, and were incubated at 45 • C for 30 min.…”