Validation of an enzyme-linked immunosorbent assay for the quantification of human IgG directed against the repeat region of the circumsporozoite protein of the parasite <it>Plasmodium falciparum</it>
<p>Abstract</p> <p>Background</p> <p>Several pre-erythrocytic malaria vaccines based on the circumsporozoite protein (CSP) antigen of <it>Plasmodium falciparum</it> are in clinical development. Vaccine immunogenicity is commonly evaluated by the determinatio...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2012-11-01
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| Series: | Malaria Journal |
| Subjects: | |
| Online Access: | http://www.malariajournal.com/content/11/1/384 |
| _version_ | 1819026168983060480 |
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| author | Clement Frederic Dewar Vincent Van Braeckel Eva Desombere Isabelle Dewerchin Marianne Swysen Christine Demoitié Marie-Ange Jongert Erik Cohen Joe Leroux-Roels Geert Cambron Pierre |
| author_facet | Clement Frederic Dewar Vincent Van Braeckel Eva Desombere Isabelle Dewerchin Marianne Swysen Christine Demoitié Marie-Ange Jongert Erik Cohen Joe Leroux-Roels Geert Cambron Pierre |
| author_sort | Clement Frederic |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Several pre-erythrocytic malaria vaccines based on the circumsporozoite protein (CSP) antigen of <it>Plasmodium falciparum</it> are in clinical development. Vaccine immunogenicity is commonly evaluated by the determination of anti-CSP antibody levels using IgG-based assays, but no standard assay is available to allow comparison of the different vaccines.</p> <p>Methods</p> <p>The validation of an anti-CSP repeat region enzyme-linked immunosorbent assay (ELISA) is described. This assay is based on the binding of serum antibodies to R32LR, a recombinant protein composed of the repeat region of <it>P. falciparum</it> CSP. In addition to the original recombinant R32LR, an easy to purify recombinant His-tagged R32LR protein has been constructed to be used as solid phase antigen in the assay. Also, hybridoma cell lines have been generated producing human anti-R32LR monoclonal antibodies to be used as a potential inexhaustible source of anti-CSP repeats standard, instead of a reference serum.</p> <p>Results</p> <p>The anti-CSP repeats ELISA was shown to be robust, specific and linear within the analytical range, and adequately fulfilled all validation criteria as defined in the ICH guidelines. Furthermore, the coefficient of variation for repeatability and intermediate precision did not exceed 23%. Non-interference was demonstrated for R32LR-binding sera, and the assay was shown to be stable over time.</p> <p>Conclusions</p> <p>This ELISA, specific for antibodies directed against the CSP repeat region, can be used as a standard assay for the determination of humoral immunogenicity in the development of any CSP-based <it>P. falciparum</it> malaria vaccine.</p> |
| first_indexed | 2024-12-21T05:22:18Z |
| format | Article |
| id | doaj.art-c38bc494612a46bc9751661add118057 |
| institution | Directory Open Access Journal |
| issn | 1475-2875 |
| language | English |
| last_indexed | 2024-12-21T05:22:18Z |
| publishDate | 2012-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Malaria Journal |
| spelling | doaj.art-c38bc494612a46bc9751661add1180572022-12-21T19:14:47ZengBMCMalaria Journal1475-28752012-11-0111138410.1186/1475-2875-11-384Validation of an enzyme-linked immunosorbent assay for the quantification of human IgG directed against the repeat region of the circumsporozoite protein of the parasite <it>Plasmodium falciparum</it>Clement FredericDewar VincentVan Braeckel EvaDesombere IsabelleDewerchin MarianneSwysen ChristineDemoitié Marie-AngeJongert ErikCohen JoeLeroux-Roels GeertCambron Pierre<p>Abstract</p> <p>Background</p> <p>Several pre-erythrocytic malaria vaccines based on the circumsporozoite protein (CSP) antigen of <it>Plasmodium falciparum</it> are in clinical development. Vaccine immunogenicity is commonly evaluated by the determination of anti-CSP antibody levels using IgG-based assays, but no standard assay is available to allow comparison of the different vaccines.</p> <p>Methods</p> <p>The validation of an anti-CSP repeat region enzyme-linked immunosorbent assay (ELISA) is described. This assay is based on the binding of serum antibodies to R32LR, a recombinant protein composed of the repeat region of <it>P. falciparum</it> CSP. In addition to the original recombinant R32LR, an easy to purify recombinant His-tagged R32LR protein has been constructed to be used as solid phase antigen in the assay. Also, hybridoma cell lines have been generated producing human anti-R32LR monoclonal antibodies to be used as a potential inexhaustible source of anti-CSP repeats standard, instead of a reference serum.</p> <p>Results</p> <p>The anti-CSP repeats ELISA was shown to be robust, specific and linear within the analytical range, and adequately fulfilled all validation criteria as defined in the ICH guidelines. Furthermore, the coefficient of variation for repeatability and intermediate precision did not exceed 23%. Non-interference was demonstrated for R32LR-binding sera, and the assay was shown to be stable over time.</p> <p>Conclusions</p> <p>This ELISA, specific for antibodies directed against the CSP repeat region, can be used as a standard assay for the determination of humoral immunogenicity in the development of any CSP-based <it>P. falciparum</it> malaria vaccine.</p>http://www.malariajournal.com/content/11/1/384Malaria<it>Plasmodium falciparum</it>Circumsporozoite proteinEnzyme-linked immunosorbent assayR32LRValidation |
| spellingShingle | Clement Frederic Dewar Vincent Van Braeckel Eva Desombere Isabelle Dewerchin Marianne Swysen Christine Demoitié Marie-Ange Jongert Erik Cohen Joe Leroux-Roels Geert Cambron Pierre Validation of an enzyme-linked immunosorbent assay for the quantification of human IgG directed against the repeat region of the circumsporozoite protein of the parasite <it>Plasmodium falciparum</it> Malaria Journal Malaria <it>Plasmodium falciparum</it> Circumsporozoite protein Enzyme-linked immunosorbent assay R32LR Validation |
| title | Validation of an enzyme-linked immunosorbent assay for the quantification of human IgG directed against the repeat region of the circumsporozoite protein of the parasite <it>Plasmodium falciparum</it> |
| title_full | Validation of an enzyme-linked immunosorbent assay for the quantification of human IgG directed against the repeat region of the circumsporozoite protein of the parasite <it>Plasmodium falciparum</it> |
| title_fullStr | Validation of an enzyme-linked immunosorbent assay for the quantification of human IgG directed against the repeat region of the circumsporozoite protein of the parasite <it>Plasmodium falciparum</it> |
| title_full_unstemmed | Validation of an enzyme-linked immunosorbent assay for the quantification of human IgG directed against the repeat region of the circumsporozoite protein of the parasite <it>Plasmodium falciparum</it> |
| title_short | Validation of an enzyme-linked immunosorbent assay for the quantification of human IgG directed against the repeat region of the circumsporozoite protein of the parasite <it>Plasmodium falciparum</it> |
| title_sort | validation of an enzyme linked immunosorbent assay for the quantification of human igg directed against the repeat region of the circumsporozoite protein of the parasite it plasmodium falciparum it |
| topic | Malaria <it>Plasmodium falciparum</it> Circumsporozoite protein Enzyme-linked immunosorbent assay R32LR Validation |
| url | http://www.malariajournal.com/content/11/1/384 |
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