Highly potent multivalent VHH antibodies against Chikungunya isolated from an alpaca naïve phage display library
Abstract Background Chikungunya virus (CHIKV) is a re-emerged mosquito-borne alphavirus that can cause musculoskeletal diseases, imposing a substantial threat to public health globally. High-affinity antibodies are need for diagnosis and treatment of CHIKV infections. As a potential diagnostic and t...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2022-05-01
|
| Series: | Journal of Nanobiotechnology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12951-022-01417-6 |
| _version_ | 1811299172652941312 |
|---|---|
| author | Qianlin Li Fuqiang Zhang Yi Lu Huan Hu Jin Wang Cheng Guo Qiang Deng Conghui Liao Qin Wu Tingsong Hu Zeliang Chen Jiahai Lu |
| author_facet | Qianlin Li Fuqiang Zhang Yi Lu Huan Hu Jin Wang Cheng Guo Qiang Deng Conghui Liao Qin Wu Tingsong Hu Zeliang Chen Jiahai Lu |
| author_sort | Qianlin Li |
| collection | DOAJ |
| description | Abstract Background Chikungunya virus (CHIKV) is a re-emerged mosquito-borne alphavirus that can cause musculoskeletal diseases, imposing a substantial threat to public health globally. High-affinity antibodies are need for diagnosis and treatment of CHIKV infections. As a potential diagnostic and therapeutic agent, the multivalent VHH antibodies is a promising tookit in nanomedicine. Here, we developed potent multivalent VHH antibodies from an alpaca naïve phage display library targeting the E2 glycoprotein of the CHIKV virus. Results In the present study, we generated 20 VHH antibodies using a naïve phage display library for binders to the CHIKV E2 glycoprotein. Of these, multivalent VHH antibodies Nb-2E8 and Nb-3C5 had specific high-affinity binding to E2 protein within the nanomolar range. The equilibrium dissociation constant (KD) was between 2.59–20.7 nM, which was 100-fold stronger than the monovalent antibodies’ affinity. Moreover, epitope mapping showed that Nb-2E8 and Nb-3C5 recognized different linear epitopes located on the E2 glycoprotein domain C and A, respectively. A facile protocol of sandwich ELISA was established using BiNb-2E8 as a capture antibody and HRP-conjugated BiNb-3C5 as a detection antibody. A good linear correlation was achieved between the OD 450 value and the E2 protein concentration in the 5–1000 ng/mL range (r = 0.9864, P < 0.0001), indicating its potential for quantitative detection of the E2 protein. Conclusions Compared to monovalent antibodies, multivalent VHH antibodies Nb-2E8 and Nb-3C5 showed high affinity and are potential candidates for diagnostic applications to better detect CHIKV virions in sera. Graphical Abstract |
| first_indexed | 2024-04-13T06:31:21Z |
| format | Article |
| id | doaj.art-be4c27b2a30f40958f318b28e39265b0 |
| institution | Directory Open Access Journal |
| issn | 1477-3155 |
| language | English |
| last_indexed | 2024-04-13T06:31:21Z |
| publishDate | 2022-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Nanobiotechnology |
| spelling | doaj.art-be4c27b2a30f40958f318b28e39265b02022-12-22T02:58:08ZengBMCJournal of Nanobiotechnology1477-31552022-05-0120111510.1186/s12951-022-01417-6Highly potent multivalent VHH antibodies against Chikungunya isolated from an alpaca naïve phage display libraryQianlin Li0Fuqiang Zhang1Yi Lu2Huan Hu3Jin Wang4Cheng Guo5Qiang Deng6Conghui Liao7Qin Wu8Tingsong Hu9Zeliang Chen10Jiahai Lu11One Health Center of Excellence for Research and Training, School of Public Health, Sun Yat-Sen UniversityCenter for Disease Control and Prevention of Southern Theater CommandHealth Effects InstituteOne Health Center of Excellence for Research and Training, School of Public Health, Sun Yat-Sen UniversityOne Health Center of Excellence for Research and Training, School of Public Health, Sun Yat-Sen UniversityCenter for Infection and Immunity, Mailman School of Public Health, Columbia UniversityOne Health Center of Excellence for Research and Training, School of Public Health, Sun Yat-Sen UniversityOne Health Center of Excellence for Research and Training, School of Public Health, Sun Yat-Sen UniversityOne Health Center of Excellence for Research and Training, School of Public Health, Sun Yat-Sen UniversityCenter for Disease Control and Prevention of Southern Theater CommandOne Health Center of Excellence for Research and Training, School of Public Health, Sun Yat-Sen UniversityOne Health Center of Excellence for Research and Training, School of Public Health, Sun Yat-Sen UniversityAbstract Background Chikungunya virus (CHIKV) is a re-emerged mosquito-borne alphavirus that can cause musculoskeletal diseases, imposing a substantial threat to public health globally. High-affinity antibodies are need for diagnosis and treatment of CHIKV infections. As a potential diagnostic and therapeutic agent, the multivalent VHH antibodies is a promising tookit in nanomedicine. Here, we developed potent multivalent VHH antibodies from an alpaca naïve phage display library targeting the E2 glycoprotein of the CHIKV virus. Results In the present study, we generated 20 VHH antibodies using a naïve phage display library for binders to the CHIKV E2 glycoprotein. Of these, multivalent VHH antibodies Nb-2E8 and Nb-3C5 had specific high-affinity binding to E2 protein within the nanomolar range. The equilibrium dissociation constant (KD) was between 2.59–20.7 nM, which was 100-fold stronger than the monovalent antibodies’ affinity. Moreover, epitope mapping showed that Nb-2E8 and Nb-3C5 recognized different linear epitopes located on the E2 glycoprotein domain C and A, respectively. A facile protocol of sandwich ELISA was established using BiNb-2E8 as a capture antibody and HRP-conjugated BiNb-3C5 as a detection antibody. A good linear correlation was achieved between the OD 450 value and the E2 protein concentration in the 5–1000 ng/mL range (r = 0.9864, P < 0.0001), indicating its potential for quantitative detection of the E2 protein. Conclusions Compared to monovalent antibodies, multivalent VHH antibodies Nb-2E8 and Nb-3C5 showed high affinity and are potential candidates for diagnostic applications to better detect CHIKV virions in sera. Graphical Abstracthttps://doi.org/10.1186/s12951-022-01417-6ChikungunyaVHH antibodyNaïve phage display libraryE2 glycoproteinEpitope |
| spellingShingle | Qianlin Li Fuqiang Zhang Yi Lu Huan Hu Jin Wang Cheng Guo Qiang Deng Conghui Liao Qin Wu Tingsong Hu Zeliang Chen Jiahai Lu Highly potent multivalent VHH antibodies against Chikungunya isolated from an alpaca naïve phage display library Journal of Nanobiotechnology Chikungunya VHH antibody Naïve phage display library E2 glycoprotein Epitope |
| title | Highly potent multivalent VHH antibodies against Chikungunya isolated from an alpaca naïve phage display library |
| title_full | Highly potent multivalent VHH antibodies against Chikungunya isolated from an alpaca naïve phage display library |
| title_fullStr | Highly potent multivalent VHH antibodies against Chikungunya isolated from an alpaca naïve phage display library |
| title_full_unstemmed | Highly potent multivalent VHH antibodies against Chikungunya isolated from an alpaca naïve phage display library |
| title_short | Highly potent multivalent VHH antibodies against Chikungunya isolated from an alpaca naïve phage display library |
| title_sort | highly potent multivalent vhh antibodies against chikungunya isolated from an alpaca naive phage display library |
| topic | Chikungunya VHH antibody Naïve phage display library E2 glycoprotein Epitope |
| url | https://doi.org/10.1186/s12951-022-01417-6 |
| work_keys_str_mv | AT qianlinli highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary AT fuqiangzhang highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary AT yilu highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary AT huanhu highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary AT jinwang highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary AT chengguo highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary AT qiangdeng highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary AT conghuiliao highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary AT qinwu highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary AT tingsonghu highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary AT zeliangchen highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary AT jiahailu highlypotentmultivalentvhhantibodiesagainstchikungunyaisolatedfromanalpacanaivephagedisplaylibrary |