Effective Cryopreservation of a Bioluminescent Auxotrophic Escherichia coli-Based Amino Acid Array to Enable Long-Term Ready-to-Use Applications
DC Field | Value | Language |
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dc.contributor.author | Ahn, Hee Tae | - |
dc.contributor.author | Jang, In Seung | - |
dc.contributor.author | Dang, Thinh Viet | - |
dc.contributor.author | Kim, Yi Hyang | - |
dc.contributor.author | Lee, Dong Hoon | - |
dc.contributor.author | Choi, Hyeun Seok | - |
dc.contributor.author | Yu, Byung Jo | - |
dc.contributor.author | Kim, Moon Il | - |
dc.date.accessioned | 2021-09-03T02:41:36Z | - |
dc.date.available | 2021-09-03T02:41:36Z | - |
dc.date.created | 2021-08-23 | - |
dc.date.issued | 2021-08 | - |
dc.identifier.issn | 2079-6374 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/82034 | - |
dc.description.abstract | Amino acid arrays comprising bioluminescent amino acid auxotrophic Escherichia coli are effective systems to quantitatively determine multiple amino acids. However, there is a need to develop a method for convenient long-term preservation of the array to enable its practical applications. Here, we reported a potential strategy to efficiently maintain cell viability within the portable array. The method involves immobilization of cells within agarose gel supplemented with an appropriate cryoprotectant in individual wells of a 96-well plate, followed by storage under freezing conditions. Six cryoprotectants, namely dimethyl sulfoxide, glycerol, ethylene glycol, polyethylene glycol, sucrose, and trehalose, were tested in the methionine (Met) auxotroph-based array. Carbohydrate-type cryoprotectants (glycerol, sucrose, and trehalose) efficiently preserved the linearity of determination of Met concentration. In particular, the array with 5% trehalose exhibited the best performance. The Met array with 5% trehalose could determine Met concentration with high linearity (R2 value = approximately 0.99) even after storage at −20◦C for up to 3 months. The clinical utilities of the Met and Leu array, preserved at −20◦C for 3 months, were also verified by successfully quantifying Met and Leu in spiked blood serum samples for the diagnosis of the corresponding metabolic diseases. This long-term preservation protocol enables the development of a ready-to-use bioluminescent E. coli-based amino acid array to quantify multiple amino acids and can replace the currently used laborious analytical methods. © 2021 by the authors. Licensee MDPI, Basel, Switzerland. | - |
dc.language | 영어 | - |
dc.language.iso | en | - |
dc.publisher | MDPI | - |
dc.relation.isPartOf | BIOSENSORS-BASEL | - |
dc.title | Effective Cryopreservation of a Bioluminescent Auxotrophic Escherichia coli-Based Amino Acid Array to Enable Long-Term Ready-to-Use Applications | - |
dc.type | Article | - |
dc.type.rims | ART | - |
dc.description.journalClass | 1 | - |
dc.identifier.wosid | 000688913000001 | - |
dc.identifier.doi | 10.3390/bios11080252 | - |
dc.identifier.bibliographicCitation | BIOSENSORS-BASEL, v.11, no.8 | - |
dc.description.isOpenAccess | N | - |
dc.identifier.scopusid | 2-s2.0-85112650705 | - |
dc.citation.title | BIOSENSORS-BASEL | - |
dc.citation.volume | 11 | - |
dc.citation.number | 8 | - |
dc.contributor.affiliatedAuthor | Ahn, Hee Tae | - |
dc.contributor.affiliatedAuthor | Dang, Thinh Viet | - |
dc.contributor.affiliatedAuthor | Lee, Dong Hoon | - |
dc.contributor.affiliatedAuthor | Kim, Moon Il | - |
dc.type.docType | Article | - |
dc.subject.keywordAuthor | Amino acid array | - |
dc.subject.keywordAuthor | Cryoprotectant | - |
dc.subject.keywordAuthor | Escherichia coli auxotroph | - |
dc.subject.keywordAuthor | Long-term preservation | - |
dc.subject.keywordAuthor | Trehalose | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
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