Protein re-surfacing of E. coli L-Asparaginase to evade pre-existing anti-drug antibodies and hypersensitivity responses
The optimal use of many biotherapeutics is restricted by Anti-drug antibodies (ADAs) and hypersensitivity responses which can affect potency and ability to administer a treatment. Here we demonstrate that Re-surfacing can be utilized as a generalizable approach to engineer proteins with extensive su...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-12-01
|
| Series: | Frontiers in Immunology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2022.1016179/full |
| _version_ | 1811209892382375936 |
|---|---|
| author | Ali Bootwala Hyun Hwan An Meghan Whitney Franklin Benjamin J. Manning Lucy Y. Xu Shruti Panchal Joseph D. Garlick Reshica Baral Michael E. Hudson Gevorg Grigoryan Mark A. Murakami Kristen Hopson Daniel S. Leventhal |
| author_facet | Ali Bootwala Hyun Hwan An Meghan Whitney Franklin Benjamin J. Manning Lucy Y. Xu Shruti Panchal Joseph D. Garlick Reshica Baral Michael E. Hudson Gevorg Grigoryan Mark A. Murakami Kristen Hopson Daniel S. Leventhal |
| author_sort | Ali Bootwala |
| collection | DOAJ |
| description | The optimal use of many biotherapeutics is restricted by Anti-drug antibodies (ADAs) and hypersensitivity responses which can affect potency and ability to administer a treatment. Here we demonstrate that Re-surfacing can be utilized as a generalizable approach to engineer proteins with extensive surface residue modifications in order to avoid binding by pre-existing ADAs. This technique was applied to E. coli Asparaginase (ASN) to produce functional mutants with up to 58 substitutions resulting in direct modification of 35% of surface residues. Re-surfaced ASNs exhibited significantly reduced binding to murine, rabbit and human polyclonal ADAs, with a negative correlation observed between binding and mutational distance from the native protein. Reductions in ADA binding correlated with diminished hypersensitivity responses in an in vivo mouse model. By using computational design approaches to traverse extended distances in mutational space while maintaining function, protein Re-surfacing may provide a means to generate novel or second line therapies for life-saving drugs with limited therapeutic alternatives. |
| first_indexed | 2024-04-12T04:46:42Z |
| format | Article |
| id | doaj.art-4a3b4c9302ed43f8bcb616f28001d45c |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-04-12T04:46:42Z |
| publishDate | 2022-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-4a3b4c9302ed43f8bcb616f28001d45c2022-12-22T03:47:27ZengFrontiers Media S.A.Frontiers in Immunology1664-32242022-12-011310.3389/fimmu.2022.10161791016179Protein re-surfacing of E. coli L-Asparaginase to evade pre-existing anti-drug antibodies and hypersensitivity responsesAli Bootwala0Hyun Hwan An1Meghan Whitney Franklin2Benjamin J. Manning3Lucy Y. Xu4Shruti Panchal5Joseph D. Garlick6Reshica Baral7Michael E. Hudson8Gevorg Grigoryan9Mark A. Murakami10Kristen Hopson11Daniel S. Leventhal12Generate Biomedicines, Somerville, MA, United StatesDepartment of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, United StatesGenerate Biomedicines, Somerville, MA, United StatesGenerate Biomedicines, Somerville, MA, United StatesGenerate Biomedicines, Somerville, MA, United StatesGenerate Biomedicines, Somerville, MA, United StatesGenerate Biomedicines, Somerville, MA, United StatesGenerate Biomedicines, Somerville, MA, United StatesGenerate Biomedicines, Somerville, MA, United StatesGenerate Biomedicines, Somerville, MA, United StatesDepartment of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, United StatesGenerate Biomedicines, Somerville, MA, United StatesGenerate Biomedicines, Somerville, MA, United StatesThe optimal use of many biotherapeutics is restricted by Anti-drug antibodies (ADAs) and hypersensitivity responses which can affect potency and ability to administer a treatment. Here we demonstrate that Re-surfacing can be utilized as a generalizable approach to engineer proteins with extensive surface residue modifications in order to avoid binding by pre-existing ADAs. This technique was applied to E. coli Asparaginase (ASN) to produce functional mutants with up to 58 substitutions resulting in direct modification of 35% of surface residues. Re-surfaced ASNs exhibited significantly reduced binding to murine, rabbit and human polyclonal ADAs, with a negative correlation observed between binding and mutational distance from the native protein. Reductions in ADA binding correlated with diminished hypersensitivity responses in an in vivo mouse model. By using computational design approaches to traverse extended distances in mutational space while maintaining function, protein Re-surfacing may provide a means to generate novel or second line therapies for life-saving drugs with limited therapeutic alternatives.https://www.frontiersin.org/articles/10.3389/fimmu.2022.1016179/fullimmunogenicityanti-drug antibodies (ADA)hypersensitivity - immunologyprotein engineeringcomputational protein designasparaginase |
| spellingShingle | Ali Bootwala Hyun Hwan An Meghan Whitney Franklin Benjamin J. Manning Lucy Y. Xu Shruti Panchal Joseph D. Garlick Reshica Baral Michael E. Hudson Gevorg Grigoryan Mark A. Murakami Kristen Hopson Daniel S. Leventhal Protein re-surfacing of E. coli L-Asparaginase to evade pre-existing anti-drug antibodies and hypersensitivity responses Frontiers in Immunology immunogenicity anti-drug antibodies (ADA) hypersensitivity - immunology protein engineering computational protein design asparaginase |
| title | Protein re-surfacing of E. coli L-Asparaginase to evade pre-existing anti-drug antibodies and hypersensitivity responses |
| title_full | Protein re-surfacing of E. coli L-Asparaginase to evade pre-existing anti-drug antibodies and hypersensitivity responses |
| title_fullStr | Protein re-surfacing of E. coli L-Asparaginase to evade pre-existing anti-drug antibodies and hypersensitivity responses |
| title_full_unstemmed | Protein re-surfacing of E. coli L-Asparaginase to evade pre-existing anti-drug antibodies and hypersensitivity responses |
| title_short | Protein re-surfacing of E. coli L-Asparaginase to evade pre-existing anti-drug antibodies and hypersensitivity responses |
| title_sort | protein re surfacing of e coli l asparaginase to evade pre existing anti drug antibodies and hypersensitivity responses |
| topic | immunogenicity anti-drug antibodies (ADA) hypersensitivity - immunology protein engineering computational protein design asparaginase |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2022.1016179/full |
| work_keys_str_mv | AT alibootwala proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT hyunhwanan proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT meghanwhitneyfranklin proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT benjaminjmanning proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT lucyyxu proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT shrutipanchal proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT josephdgarlick proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT reshicabaral proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT michaelehudson proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT gevorggrigoryan proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT markamurakami proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT kristenhopson proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses AT danielsleventhal proteinresurfacingofecolilasparaginasetoevadepreexistingantidrugantibodiesandhypersensitivityresponses |