Chapter Preparing Proteoforms of Therapeutic Proteins for Top-Down Mass Spectrometry
A characteristic of many proteoforms, derived from a single gene, is their similarity regarding the composition of atoms, making their analysis very challenging. Many overexpressed recombinant proteins are strongly associated with this problem, especially recombinant therapeutic glycoproteins from l...
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| التنسيق: | Online |
| اللغة: | الإنجليزية |
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2021
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| الوصول للمادة أونلاين: | ONIX_20210602_10.5772/intechopen.89644_439 |
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| _version_ | 1869527333932630016 |
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| author | Heikaus, Laura Schlüter, Hartmut Nurul Hidayah, Siti Gaikwad, Manasi |
| author_browse | Gaikwad, Manasi Heikaus, Laura Nurul Hidayah, Siti Schlüter, Hartmut |
| author_facet | Heikaus, Laura Schlüter, Hartmut Nurul Hidayah, Siti Gaikwad, Manasi |
| author_sort | Heikaus, Laura |
| collection | Directory of Open Access Books |
| description | A characteristic of many proteoforms, derived from a single gene, is their similarity regarding the composition of atoms, making their analysis very challenging. Many overexpressed recombinant proteins are strongly associated with this problem, especially recombinant therapeutic glycoproteins from large-scale productions. In contrast to small molecule drugs, which consist of a single defined molecule, therapeutic protein preparations are heterogenous mixtures of dozens or even hundreds of very similar species. With mass spectrometry, currently high-quality spectra of intact proteoforms can be obtained only, if the complexity of the mixture of individual proteoform-ions, entering the gas phase at the same time is low. Thus, prior to mass spectrometric analysis, an effective separation is required for getting fractions with a low number of individual proteoforms. This is especially true not only for recombinant therapeutic proteins, because of their huge heterogeneity, but also relevant for top-down proteomics. Purification of proteoforms is the bottleneck in analyzing intact proteoforms with mass spectrometry. This review is focusing on the current state of the art, especially of liquid chromatography for preparing proteoforms for mass spectrometric top-down analysis. The topic of therapeutic proteins has been chosen, because this group of proteins is most challenging regarding their proteoform analysis. |
| format | Online |
| id | doab-20.500.12854ir-70634 |
| institution | Directory of Open Access Books |
| language | eng |
| publishDate | 2021 |
| publishDateRange | 2021 |
| publishDateSort | 2021 |
| publisher | InTechOpen |
| publisherStr | InTechOpen |
| record_format | ojs |
| spelling | doab-20.500.12854ir-706342025-08-13T14:12:00Z Chapter Preparing Proteoforms of Therapeutic Proteins for Top-Down Mass Spectrometry Heikaus, Laura Schlüter, Hartmut Nurul Hidayah, Siti Gaikwad, Manasi proteoforms, top-down mass spectrometry, therapeutic proteins, liquid chromatography, protein purification parameter screening, displacement chromatography thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSB Biochemistry thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSB Biochemistry A characteristic of many proteoforms, derived from a single gene, is their similarity regarding the composition of atoms, making their analysis very challenging. Many overexpressed recombinant proteins are strongly associated with this problem, especially recombinant therapeutic glycoproteins from large-scale productions. In contrast to small molecule drugs, which consist of a single defined molecule, therapeutic protein preparations are heterogenous mixtures of dozens or even hundreds of very similar species. With mass spectrometry, currently high-quality spectra of intact proteoforms can be obtained only, if the complexity of the mixture of individual proteoform-ions, entering the gas phase at the same time is low. Thus, prior to mass spectrometric analysis, an effective separation is required for getting fractions with a low number of individual proteoforms. This is especially true not only for recombinant therapeutic proteins, because of their huge heterogeneity, but also relevant for top-down proteomics. Purification of proteoforms is the bottleneck in analyzing intact proteoforms with mass spectrometry. This review is focusing on the current state of the art, especially of liquid chromatography for preparing proteoforms for mass spectrometric top-down analysis. The topic of therapeutic proteins has been chosen, because this group of proteins is most challenging regarding their proteoform analysis. 2021-02-10T12:58:18Z 2021-06-02T10:12:13Z 2019 chapter ONIX_20210602_10.5772/intechopen.89644_439 https://library.oapen.org/handle/20.500.12657/49325 https://directory.doabooks.org/handle/20.500.12854/70634 eng open access image/jpeg image/jpeg image/jpeg n/a n/a n/a https://library.oapen.org/bitstream/20.500.12657/49325/1/69506.pdf https://library.oapen.org/bitstream/20.500.12657/49325/1/69506.pdf https://library.oapen.org/bitstream/20.500.12657/49325/1/69506.pdf InTechOpen 10.5772/intechopen.89644 10.5772/intechopen.89644 035ecc65-6737-43cf-a13a-6bdf67ce01f4 open access |
| spellingShingle | proteoforms, top-down mass spectrometry, therapeutic proteins, liquid chromatography, protein purification parameter screening, displacement chromatography thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSB Biochemistry thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSB Biochemistry Heikaus, Laura Schlüter, Hartmut Nurul Hidayah, Siti Gaikwad, Manasi Chapter Preparing Proteoforms of Therapeutic Proteins for Top-Down Mass Spectrometry |
| title | Chapter Preparing Proteoforms of Therapeutic Proteins for Top-Down Mass Spectrometry |
| title_full | Chapter Preparing Proteoforms of Therapeutic Proteins for Top-Down Mass Spectrometry |
| title_fullStr | Chapter Preparing Proteoforms of Therapeutic Proteins for Top-Down Mass Spectrometry |
| title_full_unstemmed | Chapter Preparing Proteoforms of Therapeutic Proteins for Top-Down Mass Spectrometry |
| title_short | Chapter Preparing Proteoforms of Therapeutic Proteins for Top-Down Mass Spectrometry |
| title_sort | chapter preparing proteoforms of therapeutic proteins for top down mass spectrometry |
| topic | proteoforms, top-down mass spectrometry, therapeutic proteins, liquid chromatography, protein purification parameter screening, displacement chromatography thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSB Biochemistry thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSB Biochemistry |
| topic_facet | proteoforms, top-down mass spectrometry, therapeutic proteins, liquid chromatography, protein purification parameter screening, displacement chromatography thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSB Biochemistry thema EDItEUR::P Mathematics and Science::PS Biology, life sciences::PSB Biochemistry |
| url | ONIX_20210602_10.5772/intechopen.89644_439 |
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