Purification of anti-Mycobacterium tuberculosis MPT64 immunoglobulin-Y from egg-yolk supernatant using thiophilic adsorption chromatography
The significance of immunoglobulin Y (IgY) as a particular antibody equal to mammalian IgG is well understood. However, due to a lack of reliable purification procedures, producing highly pure IgY remains problematic. In this study, we aimed to optimize the recovery of pure IgY anti-MPT64 using thio...
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HH Publisher
2024
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| Online Access: | http://psasir.upm.edu.my/id/eprint/116970/ http://psasir.upm.edu.my/id/eprint/116970/1/116970.pdf |
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| author | Kusuma, Sri Agung Fitri Fadhlillah, Muhammad Maisyarah, Intan Timur Khairunnisa, Shintani Ayunda Yusuf, Muhammad Rukayadi, Yaya Subroto, Toto |
| author_facet | Kusuma, Sri Agung Fitri Fadhlillah, Muhammad Maisyarah, Intan Timur Khairunnisa, Shintani Ayunda Yusuf, Muhammad Rukayadi, Yaya Subroto, Toto |
| author_sort | Kusuma, Sri Agung Fitri |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | The significance of immunoglobulin Y (IgY) as a particular antibody equal to mammalian IgG is well understood. However, due to a lack of reliable purification procedures, producing highly pure IgY remains problematic. In this study, we aimed to optimize the recovery of pure IgY anti-MPT64 using thiophilic adsorption chromatography. The purification of IgY anti-MPT64 was achieved by initial PEG lipid precipitation, then an optimized purification by varying the gradient concentration of elution buffer into five steps gradient (0–20, 20–40, 40–60, 60–80, and 80–100%v/v) for three injection column volume (CV) each and two steps gradient (0–50 and 50–100%v/v) in eight CV for each concentration. The obtained IgY was characterized by SDS-PAGE and dot-blot then determined its content levels using the Lowry method. The results showed that the five steps gradient purification was found to provide a better purity level of IgY than the two steps gradient. However, the IgY content obtained in the two steps gradient purification (2.2632± 0.011 mg/mL) was higher than the five steps gradient purification (1.35482 ± 0.023 mg/mL). Nevertheless, both purified IgY results can recognize MPT64 protein through a dot blot test. Therefore, it can be summarized that thiophilic adsorption chromatography with five steps gradient of purification was an efficient process to obtain a higher purity of IgY anti-MPT64, especially to be targeted as a diagnostic kit component for MPT64 detection. |
| first_indexed | 2025-11-15T14:31:40Z |
| format | Article |
| id | upm-116970 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T14:31:40Z |
| publishDate | 2024 |
| publisher | HH Publisher |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-1169702025-04-21T08:31:45Z http://psasir.upm.edu.my/id/eprint/116970/ Purification of anti-Mycobacterium tuberculosis MPT64 immunoglobulin-Y from egg-yolk supernatant using thiophilic adsorption chromatography Kusuma, Sri Agung Fitri Fadhlillah, Muhammad Maisyarah, Intan Timur Khairunnisa, Shintani Ayunda Yusuf, Muhammad Rukayadi, Yaya Subroto, Toto The significance of immunoglobulin Y (IgY) as a particular antibody equal to mammalian IgG is well understood. However, due to a lack of reliable purification procedures, producing highly pure IgY remains problematic. In this study, we aimed to optimize the recovery of pure IgY anti-MPT64 using thiophilic adsorption chromatography. The purification of IgY anti-MPT64 was achieved by initial PEG lipid precipitation, then an optimized purification by varying the gradient concentration of elution buffer into five steps gradient (0–20, 20–40, 40–60, 60–80, and 80–100%v/v) for three injection column volume (CV) each and two steps gradient (0–50 and 50–100%v/v) in eight CV for each concentration. The obtained IgY was characterized by SDS-PAGE and dot-blot then determined its content levels using the Lowry method. The results showed that the five steps gradient purification was found to provide a better purity level of IgY than the two steps gradient. However, the IgY content obtained in the two steps gradient purification (2.2632± 0.011 mg/mL) was higher than the five steps gradient purification (1.35482 ± 0.023 mg/mL). Nevertheless, both purified IgY results can recognize MPT64 protein through a dot blot test. Therefore, it can be summarized that thiophilic adsorption chromatography with five steps gradient of purification was an efficient process to obtain a higher purity of IgY anti-MPT64, especially to be targeted as a diagnostic kit component for MPT64 detection. HH Publisher 2024-03-19 Article PeerReviewed text en cc_by_nc_4 http://psasir.upm.edu.my/id/eprint/116970/1/116970.pdf Kusuma, Sri Agung Fitri and Fadhlillah, Muhammad and Maisyarah, Intan Timur and Khairunnisa, Shintani Ayunda and Yusuf, Muhammad and Rukayadi, Yaya and Subroto, Toto (2024) Purification of anti-Mycobacterium tuberculosis MPT64 immunoglobulin-Y from egg-yolk supernatant using thiophilic adsorption chromatography. Progress In Microbes and Molecular Biology, 7 (1). art. no. a0000400. pp. 1-15. ISSN 2637-1049 https://journals.hh-publisher.com/index.php/pmmb/article/view/892 10.36877/pmmb.a0000400 |
| spellingShingle | Kusuma, Sri Agung Fitri Fadhlillah, Muhammad Maisyarah, Intan Timur Khairunnisa, Shintani Ayunda Yusuf, Muhammad Rukayadi, Yaya Subroto, Toto Purification of anti-Mycobacterium tuberculosis MPT64 immunoglobulin-Y from egg-yolk supernatant using thiophilic adsorption chromatography |
| title | Purification of anti-Mycobacterium tuberculosis MPT64 immunoglobulin-Y from egg-yolk supernatant using thiophilic adsorption chromatography |
| title_full | Purification of anti-Mycobacterium tuberculosis MPT64 immunoglobulin-Y from egg-yolk supernatant using thiophilic adsorption chromatography |
| title_fullStr | Purification of anti-Mycobacterium tuberculosis MPT64 immunoglobulin-Y from egg-yolk supernatant using thiophilic adsorption chromatography |
| title_full_unstemmed | Purification of anti-Mycobacterium tuberculosis MPT64 immunoglobulin-Y from egg-yolk supernatant using thiophilic adsorption chromatography |
| title_short | Purification of anti-Mycobacterium tuberculosis MPT64 immunoglobulin-Y from egg-yolk supernatant using thiophilic adsorption chromatography |
| title_sort | purification of anti-mycobacterium tuberculosis mpt64 immunoglobulin-y from egg-yolk supernatant using thiophilic adsorption chromatography |
| url | http://psasir.upm.edu.my/id/eprint/116970/ http://psasir.upm.edu.my/id/eprint/116970/ http://psasir.upm.edu.my/id/eprint/116970/ http://psasir.upm.edu.my/id/eprint/116970/1/116970.pdf |