東華大學圖書館 |

Language: English

Help

回圖書館首頁

手機版館藏查詢

Back

Switch To: Labeled | MARC Mode | ISBD

A Semi-Automated Method to Test for ...

Selvarajan, Sabarish.

Linked to FindBook

Google Book

Amazon

博客來

A Semi-Automated Method to Test for Enzymatic Inhibition Through the Complexation of Ribonuclease a with Statistical Copolymers.

Record Type:	Electronic resources : Monograph/item
Title/Author:	A Semi-Automated Method to Test for Enzymatic Inhibition Through the Complexation of Ribonuclease a with Statistical Copolymers./
Author:	Selvarajan, Sabarish.
Published:	Ann Arbor : ProQuest Dissertations & Theses, : 2023,
Description:	128 p.
Notes:	Source: Masters Abstracts International, Volume: 84-10.
Contained By:	Masters Abstracts International84-10.
Subject:	Polymer chemistry. -
Online resource:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=30311583
ISBN:	9798379437879

A Semi-Automated Method to Test for Enzymatic Inhibition Through the Complexation of Ribonuclease a with Statistical Copolymers.
Selvarajan, Sabarish.

A Semi-Automated Method to Test for Enzymatic Inhibition Through the Complexation of Ribonuclease a with Statistical Copolymers. - Ann Arbor : ProQuest Dissertations & Theses, 2023 - 128 p.

Source: Masters Abstracts International, Volume: 84-10.

Thesis (M.S.)--Rutgers The State University of New Jersey, School of Graduate Studies, 2023.

Ribonuclease A (RNase A) is an abundant nucleic acid metabolic enzyme found in almost every eukaryotic and prokaryotic cell type that catalyzes the breakdown of RNA molecules. It is a stable and potent enzyme that hinders the handling of nucleic acids in processes such as gene therapy, gene delivery and for in-vitro diagnostics. To overcome this problem, a semi-automated platform was created to measure the capability of heteropolymers to inhibit the activity of enzyme. The proposed assay implements a linear, reporter oligonucleotide sequence (FRET donor-acceptor pair) that emits energy via fluorescence when cleaved by the enzyme. A preset polymer library was created to later be conjugated with the enzyme and the catalytic capacity was measured for each polymer-enzyme complex using the reporter oligonucleotide as the substrate. Once sufficient data was acquired, an automated data analysis method was developed to measure the enzymatic reaction constant (k) which would be used to determine the polymer's percentage inhibition, a measure of the polymer's capability to reduce RNase A's native activity, the means of which were still uncertain. This could be accomplished by destabilizing the enzyme's higher order structure, reducing the enzyme's substrate affinity at the active site, or even through incorporating steric hindrances which, although undesired, will prevent the appropriate enzyme-substrate complexing. The information obtained from the semi-automated experimentation and automated data analysis was then introduced to the pre-existing machine learning algorithm to decipher underlying patterns in terms of polymer composition and chain length. A key takeaway from this process was the importance of experimental consistency, precision, and lack of confounds. The level of confidence in the procured data plays an important role in determining how accurate the ML pipeline is at predicting subsequent generations of polymers compositions that are better suited for the intended purpose.

ISBN: 9798379437879Subjects--Topical Terms:

3173488
Polymer chemistry.
Subjects--Index Terms:

Enzymatic inhibition

A Semi-Automated Method to Test for Enzymatic Inhibition Through the Complexation of Ribonuclease a with Statistical Copolymers.
LDR:03300nmm a2200421 4500 001 2401264
005 20241022112119.5
006 m o d
007 cr#unu||||||||
008 251215s2023 ||||||||||||||||| ||eng d
020 $a 9798379437879
035 $a (MiAaPQ)AAI30311583
035 $a AAI30311583
035 $a 2401264
040 $a MiAaPQ $c MiAaPQ
100 1 $a Selvarajan, Sabarish. $3 3771345
245 1 2 $a A Semi-Automated Method to Test for Enzymatic Inhibition Through the Complexation of Ribonuclease a with Statistical Copolymers.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2023
300 $a 128 p.
500 $a Source: Masters Abstracts International, Volume: 84-10.
500 $a Advisor: Gormley, Adam J.
502 $a Thesis (M.S.)--Rutgers The State University of New Jersey, School of Graduate Studies, 2023.
520 $a Ribonuclease A (RNase A) is an abundant nucleic acid metabolic enzyme found in almost every eukaryotic and prokaryotic cell type that catalyzes the breakdown of RNA molecules. It is a stable and potent enzyme that hinders the handling of nucleic acids in processes such as gene therapy, gene delivery and for in-vitro diagnostics. To overcome this problem, a semi-automated platform was created to measure the capability of heteropolymers to inhibit the activity of enzyme. The proposed assay implements a linear, reporter oligonucleotide sequence (FRET donor-acceptor pair) that emits energy via fluorescence when cleaved by the enzyme. A preset polymer library was created to later be conjugated with the enzyme and the catalytic capacity was measured for each polymer-enzyme complex using the reporter oligonucleotide as the substrate. Once sufficient data was acquired, an automated data analysis method was developed to measure the enzymatic reaction constant (k) which would be used to determine the polymer's percentage inhibition, a measure of the polymer's capability to reduce RNase A's native activity, the means of which were still uncertain. This could be accomplished by destabilizing the enzyme's higher order structure, reducing the enzyme's substrate affinity at the active site, or even through incorporating steric hindrances which, although undesired, will prevent the appropriate enzyme-substrate complexing. The information obtained from the semi-automated experimentation and automated data analysis was then introduced to the pre-existing machine learning algorithm to decipher underlying patterns in terms of polymer composition and chain length. A key takeaway from this process was the importance of experimental consistency, precision, and lack of confounds. The level of confidence in the procured data plays an important role in determining how accurate the ML pipeline is at predicting subsequent generations of polymers compositions that are better suited for the intended purpose.
590 $a School code: 0190.
650 4 $a Polymer chemistry. $3 3173488
650 4 $a Systematic biology. $3 3173492
650 4 $a Biomedical engineering. $3 535387
650 4 $a Biochemistry. $3 518028
653 $a Enzymatic inhibition
653 $a Machine learning
653 $a NLS
653 $a PET-RAFT
653 $a Ribonuclease
653 $a Statistical copolymers
690 $a 0495
690 $a 0423
690 $a 0541
690 $a 0487
710 2 $a Rutgers The State University of New Jersey, School of Graduate Studies. $b Biomedical Engineering. $3 3685162
773 0 $t Masters Abstracts International $g 84-10.
790 $a 0190
791 $a M.S.
792 $a 2023
793 $a English
856 4 0 $u https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=30311583