The novel llama-human chimeric antibody has potent effect in lowering LDL-c levels in hPCSK9 transgenic rats
Abstract Background The advent of proprotein convertase subtilisin/kexin type 9 (PCSK9)–inhibiting drugs have provided an effective, but extremely expensive treatment for the management of low density lipoprotein (LDL). Our aim was to explore a cost-effective application of camelid anti-PCSK9 single...
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| Format: | Article |
| Language: | English |
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Wiley
2020-02-01
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| Series: | Clinical and Translational Medicine |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s40169-020-0265-2 |
| _version_ | 1818171721541222400 |
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| author | Xinyang Li Meiniang Wang Xinhua Zhang Chuxin Liu Haitao Xiang Mi Huang Yingying Ma Xiaoyan Gao Lin Jiang Xiaopan Liu Bo Li Yong Hou Xiuqing Zhang Shuang Yang Naibo Yang |
| author_facet | Xinyang Li Meiniang Wang Xinhua Zhang Chuxin Liu Haitao Xiang Mi Huang Yingying Ma Xiaoyan Gao Lin Jiang Xiaopan Liu Bo Li Yong Hou Xiuqing Zhang Shuang Yang Naibo Yang |
| author_sort | Xinyang Li |
| collection | DOAJ |
| description | Abstract Background The advent of proprotein convertase subtilisin/kexin type 9 (PCSK9)–inhibiting drugs have provided an effective, but extremely expensive treatment for the management of low density lipoprotein (LDL). Our aim was to explore a cost-effective application of camelid anti-PCSK9 single domain antibodies (sdAbs), which are high variable regions of the camelid heavy chain antibodies (VHHs), as a human PCSK9 (hPCSK9) inhibitor. One female llama was immunized with hPCSK9. Screening of high affinity anti-PCSK9 VHHs was carried out based on surface plasmon resonance (SPR) technology. We reported a lysate kinetic analysis method improving the screening efficiency. To increase the serum half-life and targeting properties, the constant region fragment of the human immunoglobulin gamma sub-type 4 (IgG4 Fc) was incorporated to form a novel llama-human chimeric molecule (VHH-hFc). Results The PCSK9 inhibiting effects of the VHH proteins were analyzed in two human liver hepatocellular cells (HepG2 and Huh7) and in the hPCSK9 transgenic Sprague–Dawley (SD) rat model. The hPCSK9 antagonistic potency of the bivalent VHH-hFc exceeded the monovalent VHH (P < 0.001) in hepatocarcinoma cells. Furthermore, the llama-human chimeric VHH-Fc protein had a similar reduction (~ 40%) of the LDL-c and total cholesterol when compared to the approved evolocumab in transgenic SD rat model, but with low cost. More surprisingly, the chimeric heavy chain antibodies could be persevered for 3 months at room temperature with little loss of the affinity. Conclusions Due to the high yield and low cost of Pichia pastoris, lipid-lowering effect and strong stability, the llama-human chimeric antibody (VHH-Fc) offers a potent therapeutic candidate for the control of the serum lipid level. |
| first_indexed | 2024-12-11T19:01:13Z |
| format | Article |
| id | doaj.art-aa55faa17fc34fee898e05381b25085c |
| institution | Directory Open Access Journal |
| issn | 2001-1326 |
| language | English |
| last_indexed | 2024-12-11T19:01:13Z |
| publishDate | 2020-02-01 |
| publisher | Wiley |
| record_format | Article |
| series | Clinical and Translational Medicine |
| spelling | doaj.art-aa55faa17fc34fee898e05381b25085c2022-12-22T00:54:00ZengWileyClinical and Translational Medicine2001-13262020-02-019111210.1186/s40169-020-0265-2The novel llama-human chimeric antibody has potent effect in lowering LDL-c levels in hPCSK9 transgenic ratsXinyang Li0Meiniang Wang1Xinhua Zhang2Chuxin Liu3Haitao Xiang4Mi Huang5Yingying Ma6Xiaoyan Gao7Lin Jiang8Xiaopan Liu9Bo Li10Yong Hou11Xiuqing Zhang12Shuang Yang13Naibo Yang14BGI Education Center, University of Chinese Academy of SciencesBGI-ShenzhenBGI-ShenzhenBGI-ShenzhenBGI-ShenzhenChina National GeneBank, BGI-ShenzhenChina National GeneBank, BGI-ShenzhenChina National GeneBank, BGI-ShenzhenBGI-ShenzhenBGI Education Center, University of Chinese Academy of SciencesBGI-ShenzhenBGI-ShenzhenBGI-ShenzhenBGI-ShenzhenBGI-ShenzhenAbstract Background The advent of proprotein convertase subtilisin/kexin type 9 (PCSK9)–inhibiting drugs have provided an effective, but extremely expensive treatment for the management of low density lipoprotein (LDL). Our aim was to explore a cost-effective application of camelid anti-PCSK9 single domain antibodies (sdAbs), which are high variable regions of the camelid heavy chain antibodies (VHHs), as a human PCSK9 (hPCSK9) inhibitor. One female llama was immunized with hPCSK9. Screening of high affinity anti-PCSK9 VHHs was carried out based on surface plasmon resonance (SPR) technology. We reported a lysate kinetic analysis method improving the screening efficiency. To increase the serum half-life and targeting properties, the constant region fragment of the human immunoglobulin gamma sub-type 4 (IgG4 Fc) was incorporated to form a novel llama-human chimeric molecule (VHH-hFc). Results The PCSK9 inhibiting effects of the VHH proteins were analyzed in two human liver hepatocellular cells (HepG2 and Huh7) and in the hPCSK9 transgenic Sprague–Dawley (SD) rat model. The hPCSK9 antagonistic potency of the bivalent VHH-hFc exceeded the monovalent VHH (P < 0.001) in hepatocarcinoma cells. Furthermore, the llama-human chimeric VHH-Fc protein had a similar reduction (~ 40%) of the LDL-c and total cholesterol when compared to the approved evolocumab in transgenic SD rat model, but with low cost. More surprisingly, the chimeric heavy chain antibodies could be persevered for 3 months at room temperature with little loss of the affinity. Conclusions Due to the high yield and low cost of Pichia pastoris, lipid-lowering effect and strong stability, the llama-human chimeric antibody (VHH-Fc) offers a potent therapeutic candidate for the control of the serum lipid level.http://link.springer.com/article/10.1186/s40169-020-0265-2PCSK9AntibodyLDL-cVHH-FcsdAbPichia pastoris |
| spellingShingle | Xinyang Li Meiniang Wang Xinhua Zhang Chuxin Liu Haitao Xiang Mi Huang Yingying Ma Xiaoyan Gao Lin Jiang Xiaopan Liu Bo Li Yong Hou Xiuqing Zhang Shuang Yang Naibo Yang The novel llama-human chimeric antibody has potent effect in lowering LDL-c levels in hPCSK9 transgenic rats Clinical and Translational Medicine PCSK9 Antibody LDL-c VHH-Fc sdAb Pichia pastoris |
| title | The novel llama-human chimeric antibody has potent effect in lowering LDL-c levels in hPCSK9 transgenic rats |
| title_full | The novel llama-human chimeric antibody has potent effect in lowering LDL-c levels in hPCSK9 transgenic rats |
| title_fullStr | The novel llama-human chimeric antibody has potent effect in lowering LDL-c levels in hPCSK9 transgenic rats |
| title_full_unstemmed | The novel llama-human chimeric antibody has potent effect in lowering LDL-c levels in hPCSK9 transgenic rats |
| title_short | The novel llama-human chimeric antibody has potent effect in lowering LDL-c levels in hPCSK9 transgenic rats |
| title_sort | novel llama human chimeric antibody has potent effect in lowering ldl c levels in hpcsk9 transgenic rats |
| topic | PCSK9 Antibody LDL-c VHH-Fc sdAb Pichia pastoris |
| url | http://link.springer.com/article/10.1186/s40169-020-0265-2 |
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