Molar masses and structure in solution of haemoglobin hyperpolymers--a common calibration of size exclusion chromatography of these artificial oxygen carriers.

We are developing artificial oxygen carriers for medical use, based on synthetic polymers--so-called hyperpolymers--obtained by cross-linking mammalian haemoglobins. One requirement with respect to the polymers is that they should not increase the oncotic pressure of blood remarkably--this can be realized by high molecular weights of the polymers with a narrow distribution. They may act as a oxygen transporting blood additive, and--in combination with a plasma expander--as a blood substitute. Another important and desired property of the artificial oxygen carrier is a low viscosity, which--first--is due to a high degree of uniformity of the polymer size (or molar mass) distribution and--sec…

A model of stepwise isovolaemic blood exchange in anaesthetised, spontaneously breathing rats to evaluate the oxygen transport efficiency of artificial oxygen carriers.

Our research pursues the production of hypo-oncotic artificial oxygen carriers, based on artificial covalently cross-linked hyperpolymeric mammalian haemoglobins. To evaluate their in vivo efficiency in oxygen delivery to the tissue we developed a small animal model of stepwise isovolaemic blood exchange in anaesthetised, spontaneously breathing rats. With the aid of a two-way respiratory micro valve for small animals the overall oxygen uptake by the tissue of the animal can be determined. Measurements of oxygen contents in arterial and mixed venous blood and of some further blood parameters together with known oxygen-binding characteristics of artificial and native oxygen carriers, permits…

Developing an artificial oxygen carrier for use in humans, we polymerize native haemoglobin and myoglobin, using bifunctional, amino group specific cross-linkers, to soluble, so-called hyperpolymers. These polymers, like other polymerized globular proteins, are members of a new class of macromolecues which consist of macromolecular base units. They all have, due to the mechanisms of the chemical reaction, broad distributions of molecular weights. Fractions of hyperpolymers of human haemoglobin were obtained by employing preparative gel-permeation (size-exclusion) chromatography. The calibration curve of analytical gel-permeation chromatography (GPC) for haemoglobin hyperpolymers was determi…

An iterative, approximative procedure is presented, to calibrate the determination of molar masses of polymers with gel-permeation chromatography, additionally using viscometry, and transformations, according to Benoit's concept of universal calibration, even if only polymers with broad molar mass distribution are available. The calculated (intermediate) values of the average molar masses from each step of iteration converge to final values. With at least two fractions of the polymers to be analyzed, and with measured intrinsic viscosities of the fractions, a calibration curve of gel-permeation chromatography and Mark-Houwink's constants can be determined. Assumptions for the use of the cal…

Divinyl Sulfone Cross-Linked Hyperpolymeric Human Haemoglobin as an Artificial Oxygen Carrier in Anaesthetized Spontaneously Breathing Rats

Hyperpolymeric haemoglobin in concentrations necessary to transport oxygen in organism to a significant extent exhibits a negligible oncotic pressure as compared to that of plasma. This property makes hyperpolymeric haemoglobins suitable for development of an artificial oxygen transporting blood additive. With such an additive - in contrast to an oxygen transporting plasma expander - combating a chronic oxygen deficit of tissue (brain, heart, kidney, extremities or in case of anaemia) is possible. Using in these cases an isoncotic oxygen carrying plasma expander instead of an additive would be even more detrimental because of fluid load to heart, at least in case of heart injury. Chronic ox…