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

Directed Evolution of Antibodies Against Complex Targets.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Directed Evolution of Antibodies Against Complex Targets./
作者:	Desai, Alec A.
出版者:	Ann Arbor : ProQuest Dissertations & Theses, : 2021,
面頁冊數:	136 p.
附註:	Source: Dissertations Abstracts International, Volume: 83-05, Section: B.
Contained By:	Dissertations Abstracts International83-05B.
標題:	Chemical engineering. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=28845321
ISBN:	9798471102453

Directed Evolution of Antibodies Against Complex Targets.
Desai, Alec A.

Directed Evolution of Antibodies Against Complex Targets. - Ann Arbor : ProQuest Dissertations & Theses, 2021 - 136 p.

Source: Dissertations Abstracts International, Volume: 83-05, Section: B.

Thesis (Ph.D.)--University of Michigan, 2021.

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

Antibodies are an emerging class of biotherapeutics that are currently used for treating a myriad of diseases, including cancer, autoimmune disorders and viral infections. Their great success in the clinic is attributable to their many attractive properties, including their high affinity, drug-like biophysical properties (high specificity, stability, solubility), long circulation times in vivo, and amenability to protein engineering for further maturing their properties. Nevertheless, there are several outstanding challenges in their generation and engineering against complex targets that we have sought to address in this work. First, we have investigated in vitro methods for systematically generating antibodies with high conformational and sequence specificity against protein aggregates that form in neurodegenerative disorders such as Alzheimer's and Parkinson's disease. We have developed novel next-generation sequencing and flow cytometry library sorting methods to identify antibodies with high specificity for aggregates associated with Alzheimer's disease (amyloid β and tau fibrils) and Parkinson's disease (α-synuclein fibrils). Our anti-amyloid Aβ antibodies rival those of the FDA-approved drug, Aducanumab, in terms of their affinity (EC50 values of ~1-10 nM) and conformational specificity while displaying much lower levels of off-target binding. We also developed a novel flow cytometry-based selection method by capturing amyloid aggregates on nanoparticles. Using this technique, we have successfully isolated conformational antibodies against tau and α-synuclein aggregates. Our tau antibodies display similar levels of affinity (EC50 values of ~0.5 nM) and conformational specificity as a leading clinical antibody, Zagotanemab, while displaying much lower levels of off-target binding. Our α-synuclein antibody shows similar affinity (EC50 value of ~10-20 nM) and substantially higher conformational specificity relative to a leading clinical-stage antibody, Cinpanemab. Moreover, we have also developed novel directed evolution methods for generating small antibodies (nanobodies) that potently neutralize SARS-CoV-2. Unexpectedly, we discovered that systematically swapping the binding loops between low affinity lead nanobodies results in unusually large increases in affinity and neutralization activity. We show that intentionally swapping nanobody binding loops during directed evolution is a powerful method for generating high-affinity nanobodies without the need for multiple rounds of affinity maturation. These approaches result in engineered antibodies and nanobodies that rival best-in-class molecules and hold great potential for advancing the field of antibody engineering to generate the next generation of safe and potent antibody drugs.

ISBN: 9798471102453Subjects--Topical Terms:

560457
Chemical engineering.
Subjects--Index Terms:

Directed evolution

Directed Evolution of Antibodies Against Complex Targets.
LDR:04006nmm a2200385 4500 001 2346286
005 20220620110507.5
008 241004s2021 ||||||||||||||||| ||eng d
020 $a 9798471102453
035 $a (MiAaPQ)AAI28845321
035 $a (MiAaPQ)umichrackham003638
035 $a AAI28845321
040 $a MiAaPQ $c MiAaPQ
100 1 $a Desai, Alec A. $3 3685365
245 1 0 $a Directed Evolution of Antibodies Against Complex Targets.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2021
300 $a 136 p.
500 $a Source: Dissertations Abstracts International, Volume: 83-05, Section: B.
500 $a Advisor: Tessier, Peter M.
502 $a Thesis (Ph.D.)--University of Michigan, 2021.
506 $a This item must not be sold to any third party vendors.
506 $a This item must not be added to any third party search indexes.
520 $a Antibodies are an emerging class of biotherapeutics that are currently used for treating a myriad of diseases, including cancer, autoimmune disorders and viral infections. Their great success in the clinic is attributable to their many attractive properties, including their high affinity, drug-like biophysical properties (high specificity, stability, solubility), long circulation times in vivo, and amenability to protein engineering for further maturing their properties. Nevertheless, there are several outstanding challenges in their generation and engineering against complex targets that we have sought to address in this work. First, we have investigated in vitro methods for systematically generating antibodies with high conformational and sequence specificity against protein aggregates that form in neurodegenerative disorders such as Alzheimer's and Parkinson's disease. We have developed novel next-generation sequencing and flow cytometry library sorting methods to identify antibodies with high specificity for aggregates associated with Alzheimer's disease (amyloid β and tau fibrils) and Parkinson's disease (α-synuclein fibrils). Our anti-amyloid Aβ antibodies rival those of the FDA-approved drug, Aducanumab, in terms of their affinity (EC50 values of ~1-10 nM) and conformational specificity while displaying much lower levels of off-target binding. We also developed a novel flow cytometry-based selection method by capturing amyloid aggregates on nanoparticles. Using this technique, we have successfully isolated conformational antibodies against tau and α-synuclein aggregates. Our tau antibodies display similar levels of affinity (EC50 values of ~0.5 nM) and conformational specificity as a leading clinical antibody, Zagotanemab, while displaying much lower levels of off-target binding. Our α-synuclein antibody shows similar affinity (EC50 value of ~10-20 nM) and substantially higher conformational specificity relative to a leading clinical-stage antibody, Cinpanemab. Moreover, we have also developed novel directed evolution methods for generating small antibodies (nanobodies) that potently neutralize SARS-CoV-2. Unexpectedly, we discovered that systematically swapping the binding loops between low affinity lead nanobodies results in unusually large increases in affinity and neutralization activity. We show that intentionally swapping nanobody binding loops during directed evolution is a powerful method for generating high-affinity nanobodies without the need for multiple rounds of affinity maturation. These approaches result in engineered antibodies and nanobodies that rival best-in-class molecules and hold great potential for advancing the field of antibody engineering to generate the next generation of safe and potent antibody drugs.
590 $a School code: 0127.
650 4 $a Chemical engineering. $3 560457
650 4 $a Microbiology. $3 536250
650 4 $a Biomedical engineering. $3 535387
650 4 $a Evolution & development. $3 3172418
653 $a Directed evolution
653 $a Antibody
653 $a Complex target
690 $a 0542
690 $a 0412
690 $a 0541
690 $a 0410
710 2 $a University of Michigan. $b Chemical Engineering. $3 2094168
773 0 $t Dissertations Abstracts International $g 83-05B.
790 $a 0127
791 $a Ph.D.
792 $a 2021
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=28845321