Highly reliable GIGA-sized synthetic human therapeutic antibody library construction
BackgroundMonoclonal antibodies (mAbs) and their derivatives are the fastest expanding category of pharmaceuticals. Efficient screening and generation of appropriate therapeutic human antibodies are important and urgent issues in the field of medicine. The successful in vitro biopanning method for a...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2023-04-01
|
| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1089395/full |
| _version_ | 1797838659849814016 |
|---|---|
| author | Chao-Yang Huang Chao-Yang Huang Ying-Yung Lok Chia-Hui Lin Szu-Liang Lai Yen-Yu Wu Chih-Yung Hu Chu-Bin Liao Chen-Hsuan Ho Yu-Ping Chou Yi-Hsuan Hsu Yu-Hsun Lo Edward Chern Edward Chern |
| author_facet | Chao-Yang Huang Chao-Yang Huang Ying-Yung Lok Chia-Hui Lin Szu-Liang Lai Yen-Yu Wu Chih-Yung Hu Chu-Bin Liao Chen-Hsuan Ho Yu-Ping Chou Yi-Hsuan Hsu Yu-Hsun Lo Edward Chern Edward Chern |
| author_sort | Chao-Yang Huang |
| collection | DOAJ |
| description | BackgroundMonoclonal antibodies (mAbs) and their derivatives are the fastest expanding category of pharmaceuticals. Efficient screening and generation of appropriate therapeutic human antibodies are important and urgent issues in the field of medicine. The successful in vitro biopanning method for antibody screening largely depends on the highly diverse, reliable and humanized CDR library. To rapidly obtain potent human antibodies, we designed and constructed a highly diverse synthetic human single-chain variable fragment (scFv) antibody library greater than a giga in size by phage display. Herein, the novel TIM-3-neutralizing antibodies with immunomodulatory functions derived from this library serve as an example to demonstrate the library’s potential for biomedical applications.MethodsThe library was designed with high stability scaffolds and six complementarity determining regions (CDRs) tailored to mimic human composition. The engineered antibody sequences were optimized for codon usage and subjected to synthesis. The six CDRs with variable length CDR-H3s were individually subjected to β-lactamase selection and then recombined for library construction. Five therapeutic target antigens were used for human antibody generation via phage library biopanning. TIM-3 antibody activity was verified by immunoactivity assays.ResultsWe have designed and constructed a highly diverse synthetic human scFv library named DSyn-1 (DCB Synthetic-1) containing 2.5 × 1010 phage clones. Three selected TIM-3-recognizing antibodies DCBT3-4, DCBT3-19, and DCBT3-22 showed significant inhibition activity by TIM-3 reporter assays at nanomolar ranges and binding affinities in sub-nanomolar ranges. Furthermore, clone DCBT3-22 was exceptionally superior with good physicochemical property and a purity of more than 98% without aggregation.ConclusionThe promising results illustrate not only the potential of the DSyn-1 library for biomedical research applications, but also the therapeutic potential of the three novel fully human TIM-3-neutralizing antibodies. |
| first_indexed | 2024-04-09T15:44:25Z |
| format | Article |
| id | doaj.art-a4b03bee94c74cbeab7e62d27c7bfb94 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-09T15:44:25Z |
| publishDate | 2023-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-a4b03bee94c74cbeab7e62d27c7bfb942023-04-27T05:19:20ZengFrontiers Media S.A.Frontiers in Immunology1664-32242023-04-011410.3389/fimmu.2023.10893951089395Highly reliable GIGA-sized synthetic human therapeutic antibody library constructionChao-Yang Huang0Chao-Yang Huang1Ying-Yung Lok2Chia-Hui Lin3Szu-Liang Lai4Yen-Yu Wu5Chih-Yung Hu6Chu-Bin Liao7Chen-Hsuan Ho8Yu-Ping Chou9Yi-Hsuan Hsu10Yu-Hsun Lo11Edward Chern12Edward Chern13niChe Lab for Stem Cell and Regenerative Medicine, Department of Biochemical Science and Technology, National Taiwan University, Taipei, TaiwanDevelopment Center for Biotechnology, New Taipei City, TaiwanDevelopment Center for Biotechnology, New Taipei City, TaiwanDevelopment Center for Biotechnology, New Taipei City, TaiwanDevelopment Center for Biotechnology, New Taipei City, TaiwanDevelopment Center for Biotechnology, New Taipei City, TaiwanDevelopment Center for Biotechnology, New Taipei City, TaiwanDevelopment Center for Biotechnology, New Taipei City, TaiwanDevelopment Center for Biotechnology, New Taipei City, TaiwanDevelopment Center for Biotechnology, New Taipei City, TaiwanDevelopment Center for Biotechnology, New Taipei City, TaiwanDevelopment Center for Biotechnology, New Taipei City, TaiwanniChe Lab for Stem Cell and Regenerative Medicine, Department of Biochemical Science and Technology, National Taiwan University, Taipei, TaiwanResearch Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, TaiwanBackgroundMonoclonal antibodies (mAbs) and their derivatives are the fastest expanding category of pharmaceuticals. Efficient screening and generation of appropriate therapeutic human antibodies are important and urgent issues in the field of medicine. The successful in vitro biopanning method for antibody screening largely depends on the highly diverse, reliable and humanized CDR library. To rapidly obtain potent human antibodies, we designed and constructed a highly diverse synthetic human single-chain variable fragment (scFv) antibody library greater than a giga in size by phage display. Herein, the novel TIM-3-neutralizing antibodies with immunomodulatory functions derived from this library serve as an example to demonstrate the library’s potential for biomedical applications.MethodsThe library was designed with high stability scaffolds and six complementarity determining regions (CDRs) tailored to mimic human composition. The engineered antibody sequences were optimized for codon usage and subjected to synthesis. The six CDRs with variable length CDR-H3s were individually subjected to β-lactamase selection and then recombined for library construction. Five therapeutic target antigens were used for human antibody generation via phage library biopanning. TIM-3 antibody activity was verified by immunoactivity assays.ResultsWe have designed and constructed a highly diverse synthetic human scFv library named DSyn-1 (DCB Synthetic-1) containing 2.5 × 1010 phage clones. Three selected TIM-3-recognizing antibodies DCBT3-4, DCBT3-19, and DCBT3-22 showed significant inhibition activity by TIM-3 reporter assays at nanomolar ranges and binding affinities in sub-nanomolar ranges. Furthermore, clone DCBT3-22 was exceptionally superior with good physicochemical property and a purity of more than 98% without aggregation.ConclusionThe promising results illustrate not only the potential of the DSyn-1 library for biomedical research applications, but also the therapeutic potential of the three novel fully human TIM-3-neutralizing antibodies.https://www.frontiersin.org/articles/10.3389/fimmu.2023.1089395/fullphage displaysingle-chain variable fragment (scFv)synthetic antibody libraryantibody therapeuticsTIM-3 |
| spellingShingle | Chao-Yang Huang Chao-Yang Huang Ying-Yung Lok Chia-Hui Lin Szu-Liang Lai Yen-Yu Wu Chih-Yung Hu Chu-Bin Liao Chen-Hsuan Ho Yu-Ping Chou Yi-Hsuan Hsu Yu-Hsun Lo Edward Chern Edward Chern Highly reliable GIGA-sized synthetic human therapeutic antibody library construction Frontiers in Immunology phage display single-chain variable fragment (scFv) synthetic antibody library antibody therapeutics TIM-3 |
| title | Highly reliable GIGA-sized synthetic human therapeutic antibody library construction |
| title_full | Highly reliable GIGA-sized synthetic human therapeutic antibody library construction |
| title_fullStr | Highly reliable GIGA-sized synthetic human therapeutic antibody library construction |
| title_full_unstemmed | Highly reliable GIGA-sized synthetic human therapeutic antibody library construction |
| title_short | Highly reliable GIGA-sized synthetic human therapeutic antibody library construction |
| title_sort | highly reliable giga sized synthetic human therapeutic antibody library construction |
| topic | phage display single-chain variable fragment (scFv) synthetic antibody library antibody therapeutics TIM-3 |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2023.1089395/full |
| work_keys_str_mv | AT chaoyanghuang highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT chaoyanghuang highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT yingyunglok highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT chiahuilin highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT szulianglai highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT yenyuwu highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT chihyunghu highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT chubinliao highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT chenhsuanho highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT yupingchou highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT yihsuanhsu highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT yuhsunlo highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT edwardchern highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction AT edwardchern highlyreliablegigasizedsynthetichumantherapeuticantibodylibraryconstruction |