VAR2CSA-specific IgG in pregnancy-associated malaria 337 part of VAR2CSA from 3D7 ( Fig. 3A ) and FCR3 (data not shown) to examine the antigen specificity of the monoclonal VSAPAM-specific IgG antibodies further. Antibodies PAM2.8, PAM3.10, PAM5.2, PAM6.1 and PAM7.5 tested positive in 3D7-VAR2CSA domain-specific ELISA ( Fig. 3A and Table 2 ), while antibodies PAM2.8, PAM3.10, PAM4.7, PAM5.2 and PAM8.1 tested positive in the FCR3-VAR2CSA ELISA (Table 2) . Control ELISA employing scrambled constructs and constructs from other PfEMP1 not implicated in the pathogenesis of PAM were consistently completely negative (data not shown). VAR2CSA constructs produced in Escherichia coli cells that should promote disulphide bond formation in secreted proteins (Barfod et al., 2006) were also consistently negative in ELISA (data not shown). Each of the VAR2CSA-reactive monoclonal antibodies had absolute specificity for either DBL3-X (PAM2.8, PAM6.1 and PAM8.1; originating from two donors) or DBL5-e 