Simultaneous Isolation and Purification of Transferrin and Immunoglobulin G from Human Serum—A New Biotech Solution
A fast and simple biotech method is presented for the simultaneous isolation and purification of transferrin (Tf) and immunoglobulin G (IgG) from the same pool-sample of human serum, yielding >98% pure proteins. Serum sample preparation was achieved by precipitation with ethacridine lactate (riva...
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| Format: | Article |
| Language: | English |
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MDPI AG
2025-02-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/30/5/993 |
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| author | Danilo Četić Goran Miljuš Zorana Dobrijević Nikola Gligorijević Aleksandra Vilotić Olgica Nedić Ana Penezić |
| author_facet | Danilo Četić Goran Miljuš Zorana Dobrijević Nikola Gligorijević Aleksandra Vilotić Olgica Nedić Ana Penezić |
| author_sort | Danilo Četić |
| collection | DOAJ |
| description | A fast and simple biotech method is presented for the simultaneous isolation and purification of transferrin (Tf) and immunoglobulin G (IgG) from the same pool-sample of human serum, yielding >98% pure proteins. Serum sample preparation was achieved by precipitation with ethacridine lactate (rivanol). Protein purification was performed with AKTA Avant 150 FPLC, using a Resource Q column. Three different buffers at pH 6.2 (MES, phosphate, and Bis-Tris) were tested. Isolated and purified proteins retained their native 3D structure, as shown by spectrofluorimetric measurements. Tf functionality was preserved, as confirmed by the retention of both the iron binding capacity and its ability to interact with the transferrin receptor (immunofluorescent staining), as well as the immunogenicity of IgG, as shown by Western blot analysis with immunodetection. The formation of IgG aggregates was avoided. This biotech method is a rapid, simple, and time-saving alternative to other methods for the isolation of extremely pure IgG and Tf, while it is also the only method so far described for their simultaneous isolation. |
| format | Article |
| id | doaj-art-714c151ce1c24829b429f09b695331ac |
| institution | DOAJ |
| issn | 1420-3049 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj-art-714c151ce1c24829b429f09b695331ac2025-08-20T02:53:22ZengMDPI AGMolecules1420-30492025-02-0130599310.3390/molecules30050993Simultaneous Isolation and Purification of Transferrin and Immunoglobulin G from Human Serum—A New Biotech SolutionDanilo Četić0Goran Miljuš1Zorana Dobrijević2Nikola Gligorijević3Aleksandra Vilotić4Olgica Nedić5Ana Penezić6Institute for the Application of Nuclear Energy INEP, University of Belgrade, Banatska 31 b, 11000 Belgrade, SerbiaInstitute for the Application of Nuclear Energy INEP, University of Belgrade, Banatska 31 b, 11000 Belgrade, SerbiaInstitute for the Application of Nuclear Energy INEP, University of Belgrade, Banatska 31 b, 11000 Belgrade, SerbiaInstitute for the Application of Nuclear Energy INEP, University of Belgrade, Banatska 31 b, 11000 Belgrade, SerbiaInstitute for the Application of Nuclear Energy INEP, University of Belgrade, Banatska 31 b, 11000 Belgrade, SerbiaInstitute for the Application of Nuclear Energy INEP, University of Belgrade, Banatska 31 b, 11000 Belgrade, SerbiaInstitute for the Application of Nuclear Energy INEP, University of Belgrade, Banatska 31 b, 11000 Belgrade, SerbiaA fast and simple biotech method is presented for the simultaneous isolation and purification of transferrin (Tf) and immunoglobulin G (IgG) from the same pool-sample of human serum, yielding >98% pure proteins. Serum sample preparation was achieved by precipitation with ethacridine lactate (rivanol). Protein purification was performed with AKTA Avant 150 FPLC, using a Resource Q column. Three different buffers at pH 6.2 (MES, phosphate, and Bis-Tris) were tested. Isolated and purified proteins retained their native 3D structure, as shown by spectrofluorimetric measurements. Tf functionality was preserved, as confirmed by the retention of both the iron binding capacity and its ability to interact with the transferrin receptor (immunofluorescent staining), as well as the immunogenicity of IgG, as shown by Western blot analysis with immunodetection. The formation of IgG aggregates was avoided. This biotech method is a rapid, simple, and time-saving alternative to other methods for the isolation of extremely pure IgG and Tf, while it is also the only method so far described for their simultaneous isolation.https://www.mdpi.com/1420-3049/30/5/993biotherapeuticsisolation and purificationion-exchangeIgGtransferrin |
| spellingShingle | Danilo Četić Goran Miljuš Zorana Dobrijević Nikola Gligorijević Aleksandra Vilotić Olgica Nedić Ana Penezić Simultaneous Isolation and Purification of Transferrin and Immunoglobulin G from Human Serum—A New Biotech Solution Molecules biotherapeutics isolation and purification ion-exchange IgG transferrin |
| title | Simultaneous Isolation and Purification of Transferrin and Immunoglobulin G from Human Serum—A New Biotech Solution |
| title_full | Simultaneous Isolation and Purification of Transferrin and Immunoglobulin G from Human Serum—A New Biotech Solution |
| title_fullStr | Simultaneous Isolation and Purification of Transferrin and Immunoglobulin G from Human Serum—A New Biotech Solution |
| title_full_unstemmed | Simultaneous Isolation and Purification of Transferrin and Immunoglobulin G from Human Serum—A New Biotech Solution |
| title_short | Simultaneous Isolation and Purification of Transferrin and Immunoglobulin G from Human Serum—A New Biotech Solution |
| title_sort | simultaneous isolation and purification of transferrin and immunoglobulin g from human serum a new biotech solution |
| topic | biotherapeutics isolation and purification ion-exchange IgG transferrin |
| url | https://www.mdpi.com/1420-3049/30/5/993 |
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