Compositions and methods for reducing ice crystal formation
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Publication Number
US-12137682-B2
Publication Date
2024-11-12
Expiration Date
Abstract
The present invention provides peptoid polymers capable of reducing or inhibiting the formation of ice crystals at sub 0° C. temperatures. Also provided are peptoid-peptide hybrids comprising the peptoid polymers provided herein. The peptoid polymers and peptoid-peptide hybrids provided herein are useful for making cryoprotectant solutions. The peptoid polymers, peptoid-peptide hybrids, and cryoprotectant solutions provided herein are useful for making antifreeze solutions, frozen food products, and cosmetic care products. Also provided herein are methods for preserving a tissue, an organ, a cell, or a biological macromolecule using the compositions described herein.
Core Innovation
The invention relates to peptoid polymer compounds and cryoprotectant solutions that inhibit ice crystal formation. The peptoid polymer has a length of from 8 to 50 monomers and comprises at least two Nhp (2-((2-hydroxypropyl)amino)acetic acid) monomers. Disclosed embodiments include peptoid and peptoid-peptide compounds, sequence-length composition distributions, alternative sequences, and compounds identified by chemical structures.
The disclosed peptoid polymers form helical structures, including polyproline-like helices, and function as cryoprotectants across temperature ranges including about 0°C to about -20°C, about -20°C to about -40°C, and about -40°C to about -200°C, with description also extending to about -196°C. The document describes cryogenic freezing and cryopreservation approaches in which biological samples or biological macromolecules are contacted with peptoid-derived cryoprotectant compositions to achieve ice crystal inhibition and improved outcomes after thawing.
In addition to general cryoprotection, the document discusses x-ray crystallography cryoprotection, including reducing crystal dissolving and/or background scattering. It further describes screening and assay approaches such as polarized light microscopy, differential scanning calorimetry and thermal hysteresis, and cell viability assays, along with examples using peptoid compounds to prevent ice formation and reduce CPA toxicity.
Claims Coverage
Independent claim coverage centers on a peptoid polymer defined by structural composition and polymer length. There are 2 main inventive features: the peptoid polymer definition and a cryoprotectant solution containing the defined peptoid polymer.
Peptoid polymer length and Nhp monomer composition
A peptoid polymer having a length of from 8 to 50 monomers, wherein the peptoid polymer comprises at least two Nhp (2-((2-hydroxypropyl) amino) acetic acid) monomers.
Cryoprotectant solution containing the peptoid polymer
A cryoprotectant solution comprising a peptoid polymer of claim 1.
Overall, the claim coverage centers on a peptoid polymer defined by monomer length (8–50) and Nhp composition (at least two), with dependent limitations tied to ice-crystal-inhibiting performance over specified temperature ranges. The coverage also includes a cryoprotectant solution that includes the defined peptoid polymer.
Stated Advantages
Reduces or inhibits ice crystal formation.
Improves post-thaw survival.
Reduces or mitigates CPA toxicity.
For x-ray crystallography cryoprotection, reduces crystal dissolving and/or background scattering.
Documented Applications
Cryoprotectant solutions for biological applications, including improved cell cryopreservation.
Cryopreservation of biological samples including tissue, organ, and cells by contacting with peptoid-derived cryoprotectant compositions.
Cryopreservation of biological macromolecules including nucleic acids, proteins, isolated proteins/protease, lipids, and liposomes by contacting with peptoid-derived cryoprotectant compositions.
Cosmetic applications and cosmetic care products.
Antifreeze and deicing applications.
Frozen-food applications and frozen food.
X-ray crystallography cryoprotection.
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