Human Antibodies Can Cross Guinea Pig Placenta and Bind Its Neonatal Fc Receptor: Implications for Studying Immune Prophylaxis and Therapy during Pregnancy
Figure 2
Expression and binding of engineered guinea pig soluble FcRn receptor. (a) This western blot confirms secretion of the guinea pig soluble FcRn by the Huh7 human liver cell line transiently transfected with pFUSE containing soluble FcRn. Shown are reduced (lane 1) and nonreduced (lane 2) guinea pig soluble receptor conjugated to Fc and blotted with rabbit anti-FCGRT polyclonal primary antibody specific for a FcRn alpha chain peptide partially conserved in the guinea pig sequence (see Figure 3(a) for the sequence). (b) Images of Coomassie stained gel (left) and a western blot demonstrating binding of the engineered guinea pig soluble FcRn receptor to a commercial human IgG column. Shown are the molecular weight markers (lane 1, with the respective mass shown), cell culture medium, column load (lane 2), column flow-through (lane 3), and column eluate (lane 4). The apparent molecular mass is lower than 50 kDa, whereas the theoretical molecular mass without glycosylation is ~43 kDa. The column binding and elution was repeated three times with similar results.
