The role of Antibody Vκ Framework 3 region towards Antigen binding: Effects on recombinant production and Protein L binding
Abstract Antibody research has traditionally focused on heavy chains, often neglecting the important complementary role of light chains in antibody formation and secretion. In the light chain, the complementarity-determining region 3 (VL-CDR3) is specifically implicated in disease states. By modulat...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2017-06-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-017-02756-3 |
| _version_ | 1818749423438528512 |
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| author | Chinh Tran-To Su Wei-Li Ling Wai-Heng Lua Jun-Jie Poh Samuel Ken-En Gan |
| author_facet | Chinh Tran-To Su Wei-Li Ling Wai-Heng Lua Jun-Jie Poh Samuel Ken-En Gan |
| author_sort | Chinh Tran-To Su |
| collection | DOAJ |
| description | Abstract Antibody research has traditionally focused on heavy chains, often neglecting the important complementary role of light chains in antibody formation and secretion. In the light chain, the complementarity-determining region 3 (VL-CDR3) is specifically implicated in disease states. By modulating VL-CDR3 exposure on the scaffold through deletions in the framework region 3 (VL-FWR3), we further investigated the effects on secretion in recombinant production and antigen binding kinetics. Our random deletions of two residues in the VL-FWR3 of a Trastuzumab model showed that the single deletions could impact recombinant production without significant effect on Her2 binding. When both the selected residues were deleted, antibody secretion was additively decreased, and so was Her2 binding kinetics. Interestingly, we also found allosteric effects on the Protein L binding site at VL-FWR1 elicited by these deletions in VL- FWR3. Together, these findings demonstrate the importance of light chain FWR3 in antigen binding, recombinant production, and antibody purification using Protein L. |
| first_indexed | 2024-12-18T04:03:33Z |
| format | Article |
| id | doaj.art-eafe5b34846144f1ab33c00235d65039 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-18T04:03:33Z |
| publishDate | 2017-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-eafe5b34846144f1ab33c00235d650392022-12-21T21:21:38ZengNature PortfolioScientific Reports2045-23222017-06-01711710.1038/s41598-017-02756-3The role of Antibody Vκ Framework 3 region towards Antigen binding: Effects on recombinant production and Protein L bindingChinh Tran-To Su0Wei-Li Ling1Wai-Heng Lua2Jun-Jie Poh3Samuel Ken-En Gan4Bioinformatics Institute, Agency for Science, Technology and Research (A*STAR)Bioinformatics Institute, Agency for Science, Technology and Research (A*STAR)Bioinformatics Institute, Agency for Science, Technology and Research (A*STAR)Bioinformatics Institute, Agency for Science, Technology and Research (A*STAR)Bioinformatics Institute, Agency for Science, Technology and Research (A*STAR)Abstract Antibody research has traditionally focused on heavy chains, often neglecting the important complementary role of light chains in antibody formation and secretion. In the light chain, the complementarity-determining region 3 (VL-CDR3) is specifically implicated in disease states. By modulating VL-CDR3 exposure on the scaffold through deletions in the framework region 3 (VL-FWR3), we further investigated the effects on secretion in recombinant production and antigen binding kinetics. Our random deletions of two residues in the VL-FWR3 of a Trastuzumab model showed that the single deletions could impact recombinant production without significant effect on Her2 binding. When both the selected residues were deleted, antibody secretion was additively decreased, and so was Her2 binding kinetics. Interestingly, we also found allosteric effects on the Protein L binding site at VL-FWR1 elicited by these deletions in VL- FWR3. Together, these findings demonstrate the importance of light chain FWR3 in antigen binding, recombinant production, and antibody purification using Protein L.https://doi.org/10.1038/s41598-017-02756-3 |
| spellingShingle | Chinh Tran-To Su Wei-Li Ling Wai-Heng Lua Jun-Jie Poh Samuel Ken-En Gan The role of Antibody Vκ Framework 3 region towards Antigen binding: Effects on recombinant production and Protein L binding Scientific Reports |
| title | The role of Antibody Vκ Framework 3 region towards Antigen binding: Effects on recombinant production and Protein L binding |
| title_full | The role of Antibody Vκ Framework 3 region towards Antigen binding: Effects on recombinant production and Protein L binding |
| title_fullStr | The role of Antibody Vκ Framework 3 region towards Antigen binding: Effects on recombinant production and Protein L binding |
| title_full_unstemmed | The role of Antibody Vκ Framework 3 region towards Antigen binding: Effects on recombinant production and Protein L binding |
| title_short | The role of Antibody Vκ Framework 3 region towards Antigen binding: Effects on recombinant production and Protein L binding |
| title_sort | role of antibody vκ framework 3 region towards antigen binding effects on recombinant production and protein l binding |
| url | https://doi.org/10.1038/s41598-017-02756-3 |
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