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 Plasmodium falciparum.
- Author
- Frédéric Clement (UGent) , Vincent Dewar, Eva Van Braeckel (UGent) , Isabelle Desombere (UGent) , Marianne Dewerchin, Christine Swysen, Marie-Ange Demoitié, Erik Jongert, Joe Cohen, Geert Leroux-Roels (UGent) and Pierre Cambron
- Organization
- Abstract
- BACKGROUND: Several pre-erythrocytic malaria vaccines based on the circumsporozoite protein (CSP) antigen of Plasmodium falciparum 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. METHODS: 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 P. falciparum 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. RESULTS: 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. CONCLUSIONS: 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 P. falciparum malaria vaccine.
- Keywords
- Plasmodium falciparum, Malaria, Circumsporozoite protein, Enzyme-linked immunosorbent assay, R32LR, Validation, CANDIDATE MALARIA VACCINE, RANDOMIZED CONTROLLED-TRIAL, IN-VITRO ASSAY, PHASE 2A TRIAL, NAIVE ADULTS, PROTECTIVE ANTIBODIES, SYNTHETIC PEPTIDES, ESCHERICHIA-COLI, EXPANDED-PROGRAM, AFRICAN CHILDREN
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Please use this url to cite or link to this publication: http://hdl.handle.net/1854/LU-3136763
- MLA
- Clement, Frédéric, et al. “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 Plasmodium Falciparum.” MALARIA JOURNAL, vol. 11, 2012, doi:10.1186/1475-2875-11-384.
- APA
- Clement, F., Dewar, V., Van Braeckel, E., Desombere, I., Dewerchin, M., Swysen, C., … Cambron, P. (2012). 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 Plasmodium falciparum. MALARIA JOURNAL, 11. https://doi.org/10.1186/1475-2875-11-384
- Chicago author-date
- Clement, Frédéric, Vincent Dewar, Eva Van Braeckel, Isabelle Desombere, Marianne Dewerchin, Christine Swysen, Marie-Ange Demoitié, et al. 2012. “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 Plasmodium Falciparum.” MALARIA JOURNAL 11. https://doi.org/10.1186/1475-2875-11-384.
- Chicago author-date (all authors)
- Clement, Frédéric, Vincent Dewar, Eva Van Braeckel, Isabelle Desombere, Marianne Dewerchin, Christine Swysen, Marie-Ange Demoitié, Erik Jongert, Joe Cohen, Geert Leroux-Roels, and Pierre Cambron. 2012. “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 Plasmodium Falciparum.” MALARIA JOURNAL 11. doi:10.1186/1475-2875-11-384.
- Vancouver
- 1.Clement F, Dewar V, Van Braeckel E, Desombere I, Dewerchin M, Swysen C, et al. 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 Plasmodium falciparum. MALARIA JOURNAL. 2012;11.
- IEEE
- [1]F. Clement et al., “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 Plasmodium falciparum.,” MALARIA JOURNAL, vol. 11, 2012.
@article{3136763, abstract = {{BACKGROUND: Several pre-erythrocytic malaria vaccines based on the circumsporozoite protein (CSP) antigen of Plasmodium falciparum 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. METHODS: 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 P. falciparum 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. RESULTS: 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. CONCLUSIONS: 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 P. falciparum malaria vaccine.}}, articleno = {{384}}, author = {{Clement, Frédéric and Dewar, Vincent and Van Braeckel, Eva and Desombere, Isabelle and Dewerchin, Marianne and Swysen, Christine and Demoitié, Marie-Ange and Jongert, Erik and Cohen, Joe and Leroux-Roels, Geert and Cambron, Pierre}}, issn = {{1475-2875}}, journal = {{MALARIA JOURNAL}}, keywords = {{Plasmodium falciparum,Malaria,Circumsporozoite protein,Enzyme-linked immunosorbent assay,R32LR,Validation,CANDIDATE MALARIA VACCINE,RANDOMIZED CONTROLLED-TRIAL,IN-VITRO ASSAY,PHASE 2A TRIAL,NAIVE ADULTS,PROTECTIVE ANTIBODIES,SYNTHETIC PEPTIDES,ESCHERICHIA-COLI,EXPANDED-PROGRAM,AFRICAN CHILDREN}}, language = {{eng}}, pages = {{15}}, 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 Plasmodium falciparum.}}, url = {{http://doi.org/10.1186/1475-2875-11-384}}, volume = {{11}}, year = {{2012}}, }
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