University of California

University of Southwestern Louisiana

John H Crowe

Lois M Crowe

Fern Tablin

Ann E Oliver

Willem F Wolkers

Nelly M Tsvetkova

Steven C Hand

John F Carpenter

Gillian M Beattie

Alberto Hayek

Ann F Oliver

Thurein M Htoo

Naomi J Walker

Lisa M Hays

Robert E Feeney

A device for holding substances during drying comprising a flask having a structure defining an opening. A pair of contiguous or juxtaposed filters is disposed in the opening. A freeze-drying assembly comprising a freeze-drying apparatus, and the device disposed in the apparatus for holding substances during freeze-drying processing. A method for processing a substance under sterile conditions comprising disposing a substance in a flask, positioning the flask in a drying apparatus, and passing a drying medium through a pair of juxtaposed filters for drying the substance.

Flask and method for drying biological materials

A dehydrated composition is provided that includes freeze-dried erythrocytic cells. Alcohol (e.g., sterol or cholesterol) is at least partially removed from erythrocytic cells including erythrocytic membranes. After removal of at least part of the alcohol, the erythrocytic cells have a low phase transition temperature range, an intermediate phase transition temperature range, and a high phase transition temperature range. The erythrocytic cells may be loaded with an oligosaccharide (e.g., trehalose) which preserves biological properties during freeze-drying and rehydration. A process for increasing cooperativity of a phase transition of an erythrocytic cell. A process for preserving and/or increasing the survival of dehydrated erythrocytic cells, including storing dehydrated erythrocytic cells having a residual water content equal to or less than about 0.30 gram of water per gram of dry weight erythrocytic cells.

Erythrocytic cells and method for preserving cells

A dehydrated composition is provided that includes freeze-dried platelets. The platelets are loaded with trehalose which preserves biological properties during freeze-drying and rehydration. The trehalose loading is conducted at a temperature of from greater than about 25&deg; C. to less than about 40&deg; C., most preferably at 37&deg; C., with the loading solution having trehalose in an amount from about 10 mM to about 50 mM. These freeze-dried platelets are substantially shelf-stable and are rehydratable so as to have a normal response to an agonist, for example, thrombin, with virtually all of the platelets participating in clot formation within about three minutes at 37&deg; C.

Therapeutic platelets and methods

Leakage from liposomes or biological cells and structural damage, which occur upon cooling through the thermotropic phase transition temperature and upon storage at temperatures below the phase transition temperature are reduced or eliminated by incorporating thermal hysteresis proteins in the liposome or cell structure. Preferred thermal hysteresis proteins are antifreeze proteins and antifreeze glycoproteins from polar fish species, and chromatographic fraction no. 8 of antifreeze glycoproteins has been found to be particularly effective.

Prevention of leakage and phase separation during thermotropic phase

The combination of trehalose and dimethyl sulfoxide is an unusually effective cryoprotectant for islets and islet-like cell clusters, as well as platelets. Islets and ICCs, when cooled through the thermotropic phase transition in the presence of this combination of treatment agents and then returned to physiologic temperature, retain their functionality. Likewise, platelets can be similarly cooled and warmed without undergoing premature activation. In general, trehalose can be incorporated into eukaryotic cells in general by suspending the cells in a trehalose solution and either cooling or warming the solution through the thermotropic transition of the cells.

Cryopreservation of human adult and fetal pancreatic cells and human

Seat mounting bracket for a boat

A method of protecting soluble proteins such that their biological activity is preserved after freezing by exposing the protein to an amino acid or trimethylamine-N-oxide and transition metal ion prior to freezing. The protected protein can then be thawed without denaturation or impairment of the protein's biological activity. The protein is preferably exposed to the amino acid or trimethylamine-N-oxide by placing it in a 25-100 mM aqueous solution of organic solute and 1 mM Zn.sup.+2. This method is especially effective in preserving the biological activity of fragile proteins such as the enzyme phosphofructokinase. The present method can be used to preserve pharmaceutically useful proteins in a frozen form for storage and distribution. The treated protein can be thawed and administered directly to a user without removing the cryoprotectant since the amino acid or oxide and trace amounts of many transition metal ions are nontoxic.

Cryogenic protection of phosphofructokinase using amino acids and zinc

A method for preserving liposomes includes freeze-drying liposomes most preferably having an average size of about 50 nm to 100 nm with a disaccharide preserving agent being present both internally (with the encapsulated liposomal contents) and externally. The disaccharide component is in a weight ratio with respect to the lipid component of from at least about 0.1:1 to not greater than about 4:1 and is preferably trehalose. When the lyophilizates are reconstituted by rehydration, the resultant liposomes can retain up to 100% of the originally encapsulated contents.

Method for preserving liposomes

A method of protecting soluble proteins such that their biological activity is preserved after freezing by exposing the protein to a carbohydrate and transition metal ion prior to freezing. The protected protein can then be thawed or lyophilized and rehydrated without denaturation of impairment of the protein's biological activity. The protein is preferably exposed to the carbohydrate by placing it in a 25-100 mM aqueous solution of carbohydrate and 2 mM Z.sup.+2. This method is especially effective in preserving the biological activity of fragile proteins such as the enzyme phosphofructokinase. The present method can be used to preserve pharmaceutically useful proteins in a frozen or freeze-dried form for storage and distribution. The treated protein can be thawed or rehydrated and administered directly to a user without removing the cryoprotectant since the carbohydrates and trace amounts of many transition metal ions are nontoxic.

Growing community of inventors

Davis, CA, United States of America

John H Crowe