Optimization of an Antibody Light Chain Framework Enhances Expression, Biophysical Properties and Pharmacokinetics
Efficacy, safety, and manufacturability of therapeutic antibodies are influenced by their biopharmaceutical and biophysical properties. These properties can be optimized by library approaches or rationale protein design. Here, we employed a protein engineering approach to modify the variable domain...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2019-09-01
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| Series: | Antibodies |
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| Online Access: | https://www.mdpi.com/2073-4468/8/3/46 |
| _version_ | 1818951151581659136 |
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| author | Patrice Douillard Michael Freissmuth Gerhard Antoine Michael Thiele Daniel Fleischanderl Peter Matthiessen Dirk Voelkel Randolf J. Kerschbaumer Friedrich Scheiflinger Nicolas Sabarth |
| author_facet | Patrice Douillard Michael Freissmuth Gerhard Antoine Michael Thiele Daniel Fleischanderl Peter Matthiessen Dirk Voelkel Randolf J. Kerschbaumer Friedrich Scheiflinger Nicolas Sabarth |
| author_sort | Patrice Douillard |
| collection | DOAJ |
| description | Efficacy, safety, and manufacturability of therapeutic antibodies are influenced by their biopharmaceutical and biophysical properties. These properties can be optimized by library approaches or rationale protein design. Here, we employed a protein engineering approach to modify the variable domain of the light chain (VL) framework of an oxidized macrophage migration inhibitory factor (oxMIF)-specific antibody. The amendment of the antibody sequence was based on homology to human germline VL genes. Three regions or positions were identified in the VL domain—L1-4, L66, L79—and mutated independently or in combination to match the closest germline V gene. None of the mutations altered oxMIF specificity or affinity, but some variants improved thermal stability, aggregation propensity, and resulted in up to five-fold higher expression. Importantly, the improved biopharmaceutical properties translated into a superior pharmacokinetic profile of the antibody. Thus, optimization of the V domain framework can ameliorate the biophysical qualities of a therapeutic antibody candidate, and as result its manufacturability, and also has the potential to improve pharmacokinetics. |
| first_indexed | 2024-12-20T09:29:56Z |
| format | Article |
| id | doaj.art-d7479ca3ea79470ea66ca2f7611c9575 |
| institution | Directory Open Access Journal |
| issn | 2073-4468 |
| language | English |
| last_indexed | 2024-12-20T09:29:56Z |
| publishDate | 2019-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibodies |
| spelling | doaj.art-d7479ca3ea79470ea66ca2f7611c95752022-12-21T19:45:06ZengMDPI AGAntibodies2073-44682019-09-01834610.3390/antib8030046antib8030046Optimization of an Antibody Light Chain Framework Enhances Expression, Biophysical Properties and PharmacokineticsPatrice Douillard0Michael Freissmuth1Gerhard Antoine2Michael Thiele3Daniel Fleischanderl4Peter Matthiessen5Dirk Voelkel6Randolf J. Kerschbaumer7Friedrich Scheiflinger8Nicolas Sabarth9Baxalta Innovations GmbH (part of Takeda), Indusriestrasse 67, 1221 Vienna, AustriaInstitute of Pharmacology, Centre of Physiology and Pharmacology, Medical University Vienna, 1090 Vienna, AustriaBaxalta Innovations GmbH (part of Takeda), Indusriestrasse 67, 1221 Vienna, AustriaBaxalta Innovations GmbH (part of Takeda), Indusriestrasse 67, 1221 Vienna, AustriaBaxalta Innovations GmbH (part of Takeda), Indusriestrasse 67, 1221 Vienna, AustriaBaxalta Innovations GmbH (part of Takeda), Indusriestrasse 67, 1221 Vienna, AustriaBaxalta Innovations GmbH (part of Takeda), Indusriestrasse 67, 1221 Vienna, AustriaBaxalta Innovations GmbH (part of Takeda), Indusriestrasse 67, 1221 Vienna, AustriaBaxalta Innovations GmbH (part of Takeda), Indusriestrasse 67, 1221 Vienna, AustriaBaxalta Innovations GmbH (part of Takeda), Indusriestrasse 67, 1221 Vienna, AustriaEfficacy, safety, and manufacturability of therapeutic antibodies are influenced by their biopharmaceutical and biophysical properties. These properties can be optimized by library approaches or rationale protein design. Here, we employed a protein engineering approach to modify the variable domain of the light chain (VL) framework of an oxidized macrophage migration inhibitory factor (oxMIF)-specific antibody. The amendment of the antibody sequence was based on homology to human germline VL genes. Three regions or positions were identified in the VL domain—L1-4, L66, L79—and mutated independently or in combination to match the closest germline V gene. None of the mutations altered oxMIF specificity or affinity, but some variants improved thermal stability, aggregation propensity, and resulted in up to five-fold higher expression. Importantly, the improved biopharmaceutical properties translated into a superior pharmacokinetic profile of the antibody. Thus, optimization of the V domain framework can ameliorate the biophysical qualities of a therapeutic antibody candidate, and as result its manufacturability, and also has the potential to improve pharmacokinetics.https://www.mdpi.com/2073-4468/8/3/46antibody engineeringthermal stabilityantibody expressionpharmacokineticsframeworkgermlineMIFaggregation |
| spellingShingle | Patrice Douillard Michael Freissmuth Gerhard Antoine Michael Thiele Daniel Fleischanderl Peter Matthiessen Dirk Voelkel Randolf J. Kerschbaumer Friedrich Scheiflinger Nicolas Sabarth Optimization of an Antibody Light Chain Framework Enhances Expression, Biophysical Properties and Pharmacokinetics Antibodies antibody engineering thermal stability antibody expression pharmacokinetics framework germline MIF aggregation |
| title | Optimization of an Antibody Light Chain Framework Enhances Expression, Biophysical Properties and Pharmacokinetics |
| title_full | Optimization of an Antibody Light Chain Framework Enhances Expression, Biophysical Properties and Pharmacokinetics |
| title_fullStr | Optimization of an Antibody Light Chain Framework Enhances Expression, Biophysical Properties and Pharmacokinetics |
| title_full_unstemmed | Optimization of an Antibody Light Chain Framework Enhances Expression, Biophysical Properties and Pharmacokinetics |
| title_short | Optimization of an Antibody Light Chain Framework Enhances Expression, Biophysical Properties and Pharmacokinetics |
| title_sort | optimization of an antibody light chain framework enhances expression biophysical properties and pharmacokinetics |
| topic | antibody engineering thermal stability antibody expression pharmacokinetics framework germline MIF aggregation |
| url | https://www.mdpi.com/2073-4468/8/3/46 |
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