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.

In-vitro haemocompatibility of dextran-protein submicron particles

“…Albumin particles (APs) and haemoglobin (HbMPs) were prepared by the one-pot procedure using 40T-and 70T-Odex as crosslinking as described earlier [23]. Briefly, equal volumes of solution 1 consisting of 0.25 M of MnCl 2 and 50 mg/mL of HSA or Hb were rapidly mixed with solution 2 containing 0.25 M of Na 2 CO 3 and 40 mg/mL of Odex (40 T or 70 T, individually) in the beaker under vigorous stirring at room temperature.…”

“…The fabrication techniques and selection of biomaterials are the fundamental steps, which can drastically impact the haemocompatibility of the particles as well as the efficacy of the proteins in the particles. Recently, we described a new promising formulation of protein submicron particles called "One-Pot formulation" [23]. The new procedure is based on the coprecipitation-crosslinking-dissolution (CCD) method where the biopolymer particles are obtained in three main steps exploiting the ability of insoluble inorganic salts to incorporate macromolecules during precipitation in aqueous solutions.…”

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]).…”