東華大學圖書館 |

語系: 繁體中文

說明(常見問題)

回圖書館首頁

手機版館藏查詢

登入

回首頁

切換: 標籤 | MARC模式 | ISBD

FindBook

Google Book

Amazon

博客來

Investigation of miRNA as Potential Biomarkers to Optimize Chinese Hamster Ovary (CHO) Process Development for Biopharmaceuticals.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Investigation of miRNA as Potential Biomarkers to Optimize Chinese Hamster Ovary (CHO) Process Development for Biopharmaceuticals./
作者:	Mullins, David.
出版者:	Ann Arbor : ProQuest Dissertations & Theses, : 2022,
面頁冊數:	112 p.
附註:	Source: Dissertations Abstracts International, Volume: 83-10, Section: B.
Contained By:	Dissertations Abstracts International83-10B.
標題:	Molecular biology. -
ISBN:	9798426809932

Investigation of miRNA as Potential Biomarkers to Optimize Chinese Hamster Ovary (CHO) Process Development for Biopharmaceuticals.
Mullins, David.

Investigation of miRNA as Potential Biomarkers to Optimize Chinese Hamster Ovary (CHO) Process Development for Biopharmaceuticals. - Ann Arbor : ProQuest Dissertations & Theses, 2022 - 112 p.

Source: Dissertations Abstracts International, Volume: 83-10, Section: B.

Thesis (Ph.D.)--University of the Sciences in Philadelphia, 2022.

This item must not be sold to any third party vendors.

Biopharmaceuticals are biologically derived drugs that are used for many clinical applications, which include vaccines, blood components, allergenics, cell and gene therapies. The development of biopharmaceuticals, known as process development, is an expensive, iterative and time-consuming activity that is required to determine the clones that produce the highest quality product as quickly and efficiently as possible. As a result, the optimization of process development has been the focus of many studies. MicroRNAs (miRNA) are small non-coding RNA that will bind mRNA targets to regulate translation. miRNA clinical biomarkers have been extensively studied as they are stable in the harsh extracellular environments of biofluids. Therefore, we hypothesized that miRNA could potentially be used as biomarkers during process development to distinguish between high- and low-production clones during process development. The investigations in this dissertation served to characterize six high- and seven low-production clones and led to the development of a miRNA panel that may be used in the early stages of process development to select high-production clones and reduce the time and cost of biopharmaceutical development. For this purpose, a novel method was initially developed to quantify intracellular titer to demonstrate the distinguishable differences in recombinant protein expression between high- and low-production clones. In the second phase of the thesis, I developed a novel spike-in experiment to evaluate normalization methods and determined that the delta delta cycle threshold (∆∆CT) method was the optimal method for the analysis of miRNA data generated from TaqMan low density array cards. Finally, the samples that were collected during the execution of an early stage of process development (Bv1) were evaluated for miRNA profile using the OpenArray microarray platform. The miRNA-mRNA interactions were identified using Ingenuity Pathway Analysis (IPA) and the entire miRNA/mRNA panel was validated with traditional quantitative real-time PCR (qPCR). While the selected panel of biomarkers could not be validated with traditional qPCR, the data set generated can provide many avenues for future directions.

ISBN: 9798426809932Subjects--Topical Terms:

517296
Molecular biology.
Subjects--Index Terms:

Biomarkers

Investigation of miRNA as Potential Biomarkers to Optimize Chinese Hamster Ovary (CHO) Process Development for Biopharmaceuticals.
LDR:03424nmm a2200373 4500 001 2349729
005 20221003074940.5
008 241004s2022 eng d
020 $a 9798426809932
035 $a (MiAaPQ)AAI29160942
035 $a AAI29160942
040 $a MiAaPQ $c MiAaPQ
100 1 $a Mullins, David. $3 780318
245 1 0 $a Investigation of miRNA as Potential Biomarkers to Optimize Chinese Hamster Ovary (CHO) Process Development for Biopharmaceuticals.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2022
300 $a 112 p.
500 $a Source: Dissertations Abstracts International, Volume: 83-10, Section: B.
500 $a Advisor: Peethambaran, Bela.
502 $a Thesis (Ph.D.)--University of the Sciences in Philadelphia, 2022.
506 $a This item must not be sold to any third party vendors.
520 $a Biopharmaceuticals are biologically derived drugs that are used for many clinical applications, which include vaccines, blood components, allergenics, cell and gene therapies. The development of biopharmaceuticals, known as process development, is an expensive, iterative and time-consuming activity that is required to determine the clones that produce the highest quality product as quickly and efficiently as possible. As a result, the optimization of process development has been the focus of many studies. MicroRNAs (miRNA) are small non-coding RNA that will bind mRNA targets to regulate translation. miRNA clinical biomarkers have been extensively studied as they are stable in the harsh extracellular environments of biofluids. Therefore, we hypothesized that miRNA could potentially be used as biomarkers during process development to distinguish between high- and low-production clones during process development. The investigations in this dissertation served to characterize six high- and seven low-production clones and led to the development of a miRNA panel that may be used in the early stages of process development to select high-production clones and reduce the time and cost of biopharmaceutical development. For this purpose, a novel method was initially developed to quantify intracellular titer to demonstrate the distinguishable differences in recombinant protein expression between high- and low-production clones. In the second phase of the thesis, I developed a novel spike-in experiment to evaluate normalization methods and determined that the delta delta cycle threshold (∆∆CT) method was the optimal method for the analysis of miRNA data generated from TaqMan low density array cards. Finally, the samples that were collected during the execution of an early stage of process development (Bv1) were evaluated for miRNA profile using the OpenArray microarray platform. The miRNA-mRNA interactions were identified using Ingenuity Pathway Analysis (IPA) and the entire miRNA/mRNA panel was validated with traditional quantitative real-time PCR (qPCR). While the selected panel of biomarkers could not be validated with traditional qPCR, the data set generated can provide many avenues for future directions.
590 $a School code: 1379.
650 4 $a Molecular biology. $3 517296
650 4 $a Cellular biology. $3 3172791
650 4 $a Bioinformatics. $3 553671
653 $a Biomarkers
653 $a Biopharmaceuticals
653 $a Chinese hamster ovary
653 $a miRNA
653 $a Process development
653 $a Protein detection
690 $a 0307
690 $a 0379
690 $a 0715
710 2 0 $a University of the Sciences in Philadelphia. $b Cell & Molecular Biology. $3 3554311
773 0 $t Dissertations Abstracts International $g 83-10B.
790 $a 1379
791 $a Ph.D.
792 $a 2022
793 $a English