Flow electroporation chamber and method

Abstract

The present invention relates to a method and apparatus for the encapsulation of substances and drugs into cells and platelets. The present invention is also related to the incorporation of thrombus dissolving drugs, such as tissue plasminogen activator and streptokinase into platelets using the apparatus described herein. The treated platelets can then be used to treat patients suffering from a thrombus blocking a blood vessel. The present invention is also related to a preparation of red blood cells that has a stable right shift of the oxygen dissociation curve.

Description

TECHNICAL FIELD The present invention relates to methods and apparatus for the encapsulation of biologically-active substances in various cell populations. More particularly, the present invention relates to a method and apparatus for the encapsulation of allosteric effectors of hemoglobin in erythrocytes by electroporation to achieve therapeutically desirable changes in the physical characteristics of the intracellular hemoglobin. BACKGROUND OF THE INVENTION In the vascular system of an adult human being, blood has a volume of about 5 to 6 liters. Approximately one half of this volume is occupied by cells, including red blood cells (erythrocytes), white blood cells (leukocytes), and blood platelets. Red blood cells comprise the majority of the cellular components of blood. Plasma, the liquid portion of blood, is approximately 90 percent water and 10 percent various solutes. These solutes include plasma proteins, organic metabolites and waste products, and inorganic compounds. Th...

AI Technical Summary

Benefits of technology

The present invention relates to a method and apparatus for the encapsulation of biologically-active substances in various cell populations. More specifically, the present invention provides an electroporation chamber that may form part of an automated, self-contained, flow apparatus for encapsulating compounds or compositions, such as inositol hexaphosphate, in red blood cells, thereby reducing the affinity of the hemoglobin for oxyge...

Problems solved by technology

Current methods of electroporation make the procedure commercially impractical on a scale suitable for commercial use.

The drawbacks associated with the liposomal technique include poor reproducibility of the IHP concentrations incorporated in the red blood cells and significant hemolysis of the red blood cells following treatment.

Additionally, commercialization is not practical because the procedure is tedious and complicated.

Treatment of the red blood cells according to the process disclosed results in a cell with unaffected activity.

The osmotic pulse technique has several shortcomings including the fact that the technique is tedious, complicated and unsuited to automation.

In addition, the results are typically unpredictable and unreliable.

For these reasons, the osmotic pulse technique has had little commercial success.

However, the results obtained by Mouneimne and his colleagues indicate that approximately 20% of the retransfused cells were lost within the first 24 hours of transfusion.

The electroporation methods disclosed in the prior art are not suitable for processing large volumes of sample, nor use of a high or repetitive electric charge.

In addition, the stability of the P50 right shift as well as the stability of the red blood cells has not proved adequate for clinical use.

Furthermore, the methods are not suitable for use in a continuous or “flow” electroporation chamber.

Continuous use of a “static” chamber results in over heating of the chamber and increased cell lysis.

Furthermore, the existing technology is unable to incorporate a sufficient quantity of IHP in a sufficient percentage of the cells being processed to dramatically change the oxygen carrying capacity of the blood.

In addition, the prior art methods require elaborate equipment and are not suited for loading red blood cells of a patient at the point of care.

Thus, the procedur...

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