Rapid and specific biotin labelling of the erythrocyte surface antigens of both cultured and ex-vivo Plasmodium parasites
Abstract Background Sensitive detection of parasite surface antigens expressed on erythrocyte membranes is necessary to further analyse the molecular pathology of malaria. This study describes a modified biotin labelling/osmotic lysis method which rapidly produces membrane extracts enriched for labe...
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ftdoajarticles:oai:doaj.org/article:73a30282bc2a457e9ec1ed42c8d66f78 2023-05-15T15:08:58+02:00 Rapid and specific biotin labelling of the erythrocyte surface antigens of both cultured and ex-vivo Plasmodium parasites Thompson Joanne Sowa Kordai MP Sharling Lisa Kyriacou Helen M Arnot David E 2007-05-01T00:00:00Z https://doi.org/10.1186/1475-2875-6-66 https://doaj.org/article/73a30282bc2a457e9ec1ed42c8d66f78 EN eng BMC http://www.malariajournal.com/content/6/1/66 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-6-66 1475-2875 https://doaj.org/article/73a30282bc2a457e9ec1ed42c8d66f78 Malaria Journal, Vol 6, Iss 1, p 66 (2007) Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 2007 ftdoajarticles https://doi.org/10.1186/1475-2875-6-66 2022-12-31T11:45:57Z Abstract Background Sensitive detection of parasite surface antigens expressed on erythrocyte membranes is necessary to further analyse the molecular pathology of malaria. This study describes a modified biotin labelling/osmotic lysis method which rapidly produces membrane extracts enriched for labelled surface antigens and also improves the efficiency of antigen recovery compared with traditional detergent extraction and surface radio-iodination. The method can also be used with ex-vivo parasites. Methods After surface labelling with biotin in the presence of the inhibitor furosemide, detergent extraction and osmotic lysis methods of enriching for the membrane fractions were compared to determine the efficiency of purification and recovery. Biotin-labelled proteins were identified on silver-stained SDS-polyacrylamide gels. Results Detergent extraction and osmotic lysis were compared for their capacity to purify biotin-labelled Plasmodium falciparum and Plasmodium chabaudi erythrocyte surface antigens. The pellet fraction formed after osmotic lysis of P. falciparum-infected erythrocytes is notably enriched in suface antigens, including PfEMP1, when compared to detergent extraction. There is also reduced co-extraction of host proteins such as spectrin and Band 3. Conclusion Biotinylation and osmotic lysis provides an improved method to label and purify parasitised erythrocyte surface antigen extracts from both in vitro and ex vivo Plasmodium parasite preparations. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Malaria Journal 6 1 66 |
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Directory of Open Access Journals: DOAJ Articles |
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ftdoajarticles |
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English |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
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Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Thompson Joanne Sowa Kordai MP Sharling Lisa Kyriacou Helen M Arnot David E Rapid and specific biotin labelling of the erythrocyte surface antigens of both cultured and ex-vivo Plasmodium parasites |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
Abstract Background Sensitive detection of parasite surface antigens expressed on erythrocyte membranes is necessary to further analyse the molecular pathology of malaria. This study describes a modified biotin labelling/osmotic lysis method which rapidly produces membrane extracts enriched for labelled surface antigens and also improves the efficiency of antigen recovery compared with traditional detergent extraction and surface radio-iodination. The method can also be used with ex-vivo parasites. Methods After surface labelling with biotin in the presence of the inhibitor furosemide, detergent extraction and osmotic lysis methods of enriching for the membrane fractions were compared to determine the efficiency of purification and recovery. Biotin-labelled proteins were identified on silver-stained SDS-polyacrylamide gels. Results Detergent extraction and osmotic lysis were compared for their capacity to purify biotin-labelled Plasmodium falciparum and Plasmodium chabaudi erythrocyte surface antigens. The pellet fraction formed after osmotic lysis of P. falciparum-infected erythrocytes is notably enriched in suface antigens, including PfEMP1, when compared to detergent extraction. There is also reduced co-extraction of host proteins such as spectrin and Band 3. Conclusion Biotinylation and osmotic lysis provides an improved method to label and purify parasitised erythrocyte surface antigen extracts from both in vitro and ex vivo Plasmodium parasite preparations. |
format |
Article in Journal/Newspaper |
author |
Thompson Joanne Sowa Kordai MP Sharling Lisa Kyriacou Helen M Arnot David E |
author_facet |
Thompson Joanne Sowa Kordai MP Sharling Lisa Kyriacou Helen M Arnot David E |
author_sort |
Thompson Joanne |
title |
Rapid and specific biotin labelling of the erythrocyte surface antigens of both cultured and ex-vivo Plasmodium parasites |
title_short |
Rapid and specific biotin labelling of the erythrocyte surface antigens of both cultured and ex-vivo Plasmodium parasites |
title_full |
Rapid and specific biotin labelling of the erythrocyte surface antigens of both cultured and ex-vivo Plasmodium parasites |
title_fullStr |
Rapid and specific biotin labelling of the erythrocyte surface antigens of both cultured and ex-vivo Plasmodium parasites |
title_full_unstemmed |
Rapid and specific biotin labelling of the erythrocyte surface antigens of both cultured and ex-vivo Plasmodium parasites |
title_sort |
rapid and specific biotin labelling of the erythrocyte surface antigens of both cultured and ex-vivo plasmodium parasites |
publisher |
BMC |
publishDate |
2007 |
url |
https://doi.org/10.1186/1475-2875-6-66 https://doaj.org/article/73a30282bc2a457e9ec1ed42c8d66f78 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Malaria Journal, Vol 6, Iss 1, p 66 (2007) |
op_relation |
http://www.malariajournal.com/content/6/1/66 https://doaj.org/toc/1475-2875 doi:10.1186/1475-2875-6-66 1475-2875 https://doaj.org/article/73a30282bc2a457e9ec1ed42c8d66f78 |
op_doi |
https://doi.org/10.1186/1475-2875-6-66 |
container_title |
Malaria Journal |
container_volume |
6 |
container_issue |
1 |
container_start_page |
66 |
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1766340225888419840 |