Simple paired heavy- and light-chain antibody repertoire sequencing using endoplasmic reticulum microsomes
Abstract Existing methods for paired antibody heavy- and light-chain repertoire sequencing rely on specialized equipment and are limited by their commercial availability and high costs. Here, we report a novel simple and cost-effective emulsion-based single-cell paired antibody repertoire sequencing...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2018-04-01
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| Series: | Genome Medicine |
| Online Access: | http://link.springer.com/article/10.1186/s13073-018-0542-5 |
| _version_ | 1818307653317689344 |
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| author | Praneeth Reddy Devulapally Jörg Bürger Thorsten Mielke Zoltán Konthur Hans Lehrach Marie-Laure Yaspo Jörn Glökler Hans-Jörg Warnatz |
| author_facet | Praneeth Reddy Devulapally Jörg Bürger Thorsten Mielke Zoltán Konthur Hans Lehrach Marie-Laure Yaspo Jörn Glökler Hans-Jörg Warnatz |
| author_sort | Praneeth Reddy Devulapally |
| collection | DOAJ |
| description | Abstract Existing methods for paired antibody heavy- and light-chain repertoire sequencing rely on specialized equipment and are limited by their commercial availability and high costs. Here, we report a novel simple and cost-effective emulsion-based single-cell paired antibody repertoire sequencing method that employs only basic laboratory equipment. We performed a proof-of-concept using mixed mouse hybridoma cells and we also showed that our method can be used for discovery of novel antigen-specific monoclonal antibodies by sequencing human CD19+ B cell IgM and IgG repertoires isolated from peripheral whole blood before and seven days after Td (Tetanus toxoid/Diphtheria toxoid) booster immunization. We anticipate broad applicability of our method for providing insights into adaptive immune responses associated with various diseases, vaccinations, and cancer immunotherapies. |
| first_indexed | 2024-12-13T07:01:48Z |
| format | Article |
| id | doaj.art-6f45a287fbae4eae96a68e9f95f85cca |
| institution | Directory Open Access Journal |
| issn | 1756-994X |
| language | English |
| last_indexed | 2024-12-13T07:01:48Z |
| publishDate | 2018-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Genome Medicine |
| spelling | doaj.art-6f45a287fbae4eae96a68e9f95f85cca2022-12-21T23:55:55ZengBMCGenome Medicine1756-994X2018-04-0110111210.1186/s13073-018-0542-5Simple paired heavy- and light-chain antibody repertoire sequencing using endoplasmic reticulum microsomesPraneeth Reddy Devulapally0Jörg Bürger1Thorsten Mielke2Zoltán Konthur3Hans Lehrach4Marie-Laure Yaspo5Jörn Glökler6Hans-Jörg Warnatz7Otto Warburg Laboratory Gene Regulation and Systems Biology of Cancer, Max Planck Institute for Molecular GeneticsMicroscopy and Cryo-Electron Microscopy Service Group, Max Planck Institute for Molecular GeneticsMicroscopy and Cryo-Electron Microscopy Service Group, Max Planck Institute for Molecular GeneticsDepartment of Biomolecular Systems, Max Planck Institute of Colloids and InterfacesAlacris Theranostics GmbHOtto Warburg Laboratory Gene Regulation and Systems Biology of Cancer, Max Planck Institute for Molecular GeneticsDepartment of Molecular Biotechnology and Functional Genomics, Institute of Applied Biosciences, Technical University of Applied Sciences WildauOtto Warburg Laboratory Gene Regulation and Systems Biology of Cancer, Max Planck Institute for Molecular GeneticsAbstract Existing methods for paired antibody heavy- and light-chain repertoire sequencing rely on specialized equipment and are limited by their commercial availability and high costs. Here, we report a novel simple and cost-effective emulsion-based single-cell paired antibody repertoire sequencing method that employs only basic laboratory equipment. We performed a proof-of-concept using mixed mouse hybridoma cells and we also showed that our method can be used for discovery of novel antigen-specific monoclonal antibodies by sequencing human CD19+ B cell IgM and IgG repertoires isolated from peripheral whole blood before and seven days after Td (Tetanus toxoid/Diphtheria toxoid) booster immunization. We anticipate broad applicability of our method for providing insights into adaptive immune responses associated with various diseases, vaccinations, and cancer immunotherapies.http://link.springer.com/article/10.1186/s13073-018-0542-5 |
| spellingShingle | Praneeth Reddy Devulapally Jörg Bürger Thorsten Mielke Zoltán Konthur Hans Lehrach Marie-Laure Yaspo Jörn Glökler Hans-Jörg Warnatz Simple paired heavy- and light-chain antibody repertoire sequencing using endoplasmic reticulum microsomes Genome Medicine |
| title | Simple paired heavy- and light-chain antibody repertoire sequencing using endoplasmic reticulum microsomes |
| title_full | Simple paired heavy- and light-chain antibody repertoire sequencing using endoplasmic reticulum microsomes |
| title_fullStr | Simple paired heavy- and light-chain antibody repertoire sequencing using endoplasmic reticulum microsomes |
| title_full_unstemmed | Simple paired heavy- and light-chain antibody repertoire sequencing using endoplasmic reticulum microsomes |
| title_short | Simple paired heavy- and light-chain antibody repertoire sequencing using endoplasmic reticulum microsomes |
| title_sort | simple paired heavy and light chain antibody repertoire sequencing using endoplasmic reticulum microsomes |
| url | http://link.springer.com/article/10.1186/s13073-018-0542-5 |
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