# US 11,998,002 B2 · Cryopreserving processes

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- Number: US 11,998,002 B2
- Document ID: US11998002B2
- Status: Granted
- Kind: B2
- Application number: 16/981,421
- Filed: 2019-03-15
- Published: 2021-01-14
- Granted: 2024-06-04
- Earliest claimed: 2018-03-16
- Assignees: University of Warwick
- Applicants: University of Warwick
- Inventors: Matthew Gibson; Christopher Stubbs; Trisha Bailey
- Web page: https://patentgrep.com/patent/US11998002B2

## CPC

- A01N 1/125: Freeze protecting agents, e.g. cryoprotectants or osmolarity regulators
- A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS

## Abstract

The present invention relates to processes for producing compositions for the cryopreservation of biological materials, e.g. cells and proteins. The compositions comprise a polyampholyte polymer. The invention also provides certain cryopreserving compositions comprising the polyampholyte polymer.

## Claims

### Claim 1

A process for producing a cryopreserved composition comprising biological material, the process comprising the step:
(a) freezing biological material at a cryopreserving temperature in a cryopreserving composition comprising a compound of Formula I:
wherein n is 20-5000,
wherein the biological material is selected from the group consisting of cells, monolayers of cells and spheroids.

### Claim 2 (depends on 1)

The process as claimed in claim 1, wherein n is 400-600.

### Claim 3 (depends on 1)

The process as claimed in claim 1, wherein the final concentration of the compound of Formula I in the cryopreserving composition is 1-200 mg/ml, 1-40 mg/ml, or 25-150 mg/ml.

### Claim 4 (depends on 1)

The process as claimed in claim 1, wherein the cryopreserving composition additionally comprises one or more components selected from the group consisting of an aqueous buffer, an antibiotic, a sugar, an anticoagulant, an antioxidant, a pH indicator, glycerol and DMSO.

### Claim 5 (depends on 4)

The process as claimed in claim 4, wherein the cryopreserving composition comprises 2-5% DMSO.

### Claim 6 (depends on 1)

The process as claimed in claim 1, wherein the cells are red blood cells or nucleated cells.

### Claim 7 (depends on 1)

A process for producing a biological material, the process comprising the steps:
(a) thawing a frozen cryopreserving composition comprising a compound of Formula I as defined in claim 1 and biological material, wherein the biological material is selected from the group consisting of cells, monolayers of cells and spheroids; and optionally
(b) removing and/or isolating the biological material from the thawed composition.

### Claim 8

A cryopreserving composition comprising a compound of Formula I,
wherein n is 20-5000,
wherein the concentration of the compound of Formula I in the composition is 1-15 mg/ml, 25-150 mg/ml, 50-200 mg/ml, 75-150 mg/ml, or about 100 mg/ml.

### Claim 9 (depends on 8)

The cryopreserving composition as claimed in claim 8, wherein the composition does not comprise both KI and KBr.

### Claim 10 (depends on 8)

The cryopreserving composition as claimed in claim 8, wherein the composition additionally comprises biological material, wherein the biological material is selected from the group consisting of cells, monolayers of cells and spheroids.

### Claim 11

A cryopreserving composition comprising a compound of Formula I
wherein n is 20-5000,
wherein the cryopreserving composition additionally comprises:
(a) a physiologically-acceptable buffer; or
(b) biological material, wherein the biological material is selected from the group consisting of cells, monolayers of cells and spheroids.

### Claim 12 (depends on 11)

The cryopreserving composition as claimed in claim 11, wherein the cryopreserving composition is in a frozen state.

### Claim 13

A kit comprising:
(a) a compound of Formula I,
wherein n is 20-5000,
together with
(b) one or more components selected from the group consisting of an aqueous buffer, an antibiotic, a sugar, an anticoagulant, an antioxidant, a pH indicator, glycerol and DMSO.