Protein photodegradation in the visible range? Insights into protein photooxidation with respect to protein concentration
Visible light (400–800 nm) can lead to photooxidation of protein formulations, which might impair protein integrity. However, the relevant mechanism of photooxidation upon visible light exposure is still unclear for therapeutic proteins, since proteinogenic structures do not absorb light in the visi...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-12-01
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| Series: | International Journal of Pharmaceutics: X |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590156722000469 |
| _version_ | 1797798673531273216 |
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| author | Elena Hipper Florian Lehmann Wolfgang Kaiser Göran Hübner Julia Buske Michaela Blech Dariush Hinderberger Patrick Garidel |
| author_facet | Elena Hipper Florian Lehmann Wolfgang Kaiser Göran Hübner Julia Buske Michaela Blech Dariush Hinderberger Patrick Garidel |
| author_sort | Elena Hipper |
| collection | DOAJ |
| description | Visible light (400–800 nm) can lead to photooxidation of protein formulations, which might impair protein integrity. However, the relevant mechanism of photooxidation upon visible light exposure is still unclear for therapeutic proteins, since proteinogenic structures do not absorb light in the visible range. Here, we show that exposure of monoclonal antibody formulations to visible light, lead to the formation of reactive oxygen species (ROS), which subsequently induce specific protein degradations. The formation of ROS and singlet oxygen upon visible light exposure is investigated using electron paramagnetic resonance (EPR) spectroscopy. We describe the initial formation of ROS, most likely after direct reaction of molecular oxygen with a triplet state photosensitizer, generated from intersystem crossing of the excited singlet state. Since these radicals affect the oxygen content in the headspace of the vial, we monitored photooxidation of these mAb formulations. With increasing protein concentrations, we found (i) a decreasing headspace oxygen content in the sample, (ii) a higher relative number of radicals in solution and (iii) a higher protein degradation. Thus, the protein concentration dependence indicates the presence of higher concentration of a currently unknown photosensitizer. |
| first_indexed | 2024-03-13T04:07:24Z |
| format | Article |
| id | doaj.art-7cc94d0fe43b4541ae395f6342387dc1 |
| institution | Directory Open Access Journal |
| issn | 2590-1567 |
| language | English |
| last_indexed | 2024-03-13T04:07:24Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | International Journal of Pharmaceutics: X |
| spelling | doaj.art-7cc94d0fe43b4541ae395f6342387dc12023-06-21T06:59:09ZengElsevierInternational Journal of Pharmaceutics: X2590-15672023-12-015100155Protein photodegradation in the visible range? Insights into protein photooxidation with respect to protein concentrationElena Hipper0Florian Lehmann1Wolfgang Kaiser2Göran Hübner3Julia Buske4Michaela Blech5Dariush Hinderberger6Patrick Garidel7Institute of Chemistry, Martin-Luther-Universität Halle-Wittenberg, von-Danckelmann-Platz 4, 06120 Halle, GermanyInstitute of Chemistry, Martin-Luther-Universität Halle-Wittenberg, von-Danckelmann-Platz 4, 06120 Halle, GermanyBoehringer Ingelheim Pharma GmbH & Co. KG, Innovation Unit, PDB-TIP, Birkendorfer Strasse 65, 88397 Biberach an der Riss, GermanyBoehringer Ingelheim Pharma GmbH & Co. KG, Innovation Unit, ADB, Birkendorfer Strasse 65, 88397 Biberach an der Riss, GermanyBoehringer Ingelheim Pharma GmbH & Co. KG, Innovation Unit, PDB-TIP, Birkendorfer Strasse 65, 88397 Biberach an der Riss, GermanyBoehringer Ingelheim Pharma GmbH & Co. KG, Innovation Unit, PDB-TIP, Birkendorfer Strasse 65, 88397 Biberach an der Riss, GermanyInstitute of Chemistry, Martin-Luther-Universität Halle-Wittenberg, von-Danckelmann-Platz 4, 06120 Halle, GermanyInstitute of Chemistry, Martin-Luther-Universität Halle-Wittenberg, von-Danckelmann-Platz 4, 06120 Halle, Germany; Boehringer Ingelheim Pharma GmbH & Co. KG, Innovation Unit, PDB-TIP, Birkendorfer Strasse 65, 88397 Biberach an der Riss, Germany; Corresponding author at: Institute of Chemistry, Martin-Luther-Universität Halle-Wittenberg, von-Danckelmann-Platz 4, 06120 Halle, Germany; Boehringer Ingelheim Pharma GmbH & Co. KG, Innovation Unit, PDB-TIP, Birkendorfer Strasse 65, 88397 Biberach an der Riss, Germany.Visible light (400–800 nm) can lead to photooxidation of protein formulations, which might impair protein integrity. However, the relevant mechanism of photooxidation upon visible light exposure is still unclear for therapeutic proteins, since proteinogenic structures do not absorb light in the visible range. Here, we show that exposure of monoclonal antibody formulations to visible light, lead to the formation of reactive oxygen species (ROS), which subsequently induce specific protein degradations. The formation of ROS and singlet oxygen upon visible light exposure is investigated using electron paramagnetic resonance (EPR) spectroscopy. We describe the initial formation of ROS, most likely after direct reaction of molecular oxygen with a triplet state photosensitizer, generated from intersystem crossing of the excited singlet state. Since these radicals affect the oxygen content in the headspace of the vial, we monitored photooxidation of these mAb formulations. With increasing protein concentrations, we found (i) a decreasing headspace oxygen content in the sample, (ii) a higher relative number of radicals in solution and (iii) a higher protein degradation. Thus, the protein concentration dependence indicates the presence of higher concentration of a currently unknown photosensitizer.http://www.sciencedirect.com/science/article/pii/S2590156722000469Monoclonal antibodyVisible light exposurePhotooxidationPolysorbate 20PS20Electron paramagnetic resonance |
| spellingShingle | Elena Hipper Florian Lehmann Wolfgang Kaiser Göran Hübner Julia Buske Michaela Blech Dariush Hinderberger Patrick Garidel Protein photodegradation in the visible range? Insights into protein photooxidation with respect to protein concentration International Journal of Pharmaceutics: X Monoclonal antibody Visible light exposure Photooxidation Polysorbate 20 PS20 Electron paramagnetic resonance |
| title | Protein photodegradation in the visible range? Insights into protein photooxidation with respect to protein concentration |
| title_full | Protein photodegradation in the visible range? Insights into protein photooxidation with respect to protein concentration |
| title_fullStr | Protein photodegradation in the visible range? Insights into protein photooxidation with respect to protein concentration |
| title_full_unstemmed | Protein photodegradation in the visible range? Insights into protein photooxidation with respect to protein concentration |
| title_short | Protein photodegradation in the visible range? Insights into protein photooxidation with respect to protein concentration |
| title_sort | protein photodegradation in the visible range insights into protein photooxidation with respect to protein concentration |
| topic | Monoclonal antibody Visible light exposure Photooxidation Polysorbate 20 PS20 Electron paramagnetic resonance |
| url | http://www.sciencedirect.com/science/article/pii/S2590156722000469 |
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