Optimizing Malaria Antigen Quantification

The MAP assay was previously optimized for quantification of Ab to multiple malarial antigens [30] (link), including VAR2CSA sequences [24] (link). To determine the optimal concentration of ID1-ID2a, different amounts of the ID1-ID2a recombinant proteins were coupled to 1 million microspheres (SeroMAP beads, Luminex Corp., Austin, TX), including 1.6, 5, and 10 µg of the 3D7 protein and 1, 5, 10, and 15 µg of the FCR3 protein. ID1-ID2 (3D7) at 1.6 µg/million microspheres and ID1-ID2 (FCR3) at 5 µg/million microspheres were found to be optimal. DBL1 (3D7 and 7G8), DBL1+2 FCR3, DBL2 FCR3, DBL3 (FCR3) and DBL5 (FCR3) were coupled at 1 µg/million microspheres [24] (link); FV2 was coupled at 3 µg protein/million microspheres based on previous optimization studies [23] (link).

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Babakhanyan A., Leke R.G., Salanti A., Bobbili N., Gwanmesia P., Leke R.J., Quakyi I.A., Chen J.J, & Taylor D.W. (2014). The Antibody Response of Pregnant Cameroonian Women to VAR2CSA ID1-ID2a, a Small Recombinant Protein Containing the CSA-Binding Site. PLoS ONE, 9(2), e88173.

Publication 2014

Antigens Assay Austin Malarial Microspheres Protein Recombinant proteins

Corresponding Organization : University of Hawaiʻi at Mānoa

Other organizations : Université de Yaoundé I, University of Copenhagen, University of Ghana

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Variable analysis

independent variables

Concentration of ID1-ID2a recombinant proteins coupled to microspheres (1.6, 5, and 10 µg of the 3D7 protein, and 1, 5, 10, and 15 µg of the FCR3 protein)
Concentration of other antigens coupled to microspheres (DBL1 (3D7 and 7G8), DBL1+2 FCR3, DBL2 FCR3, DBL3 (FCR3) and DBL5 (FCR3) at 1 µg/million microspheres, FV2 at 3 µg protein/million microspheres)

dependent variables

Optimal concentration of ID1-ID2a recombinant proteins for coupling to microspheres

control variables

Number of microspheres (1 million)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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