Solid Phase-Based Cross-Matching as Solution for Kidney Allograft Recipients Pretreated with Therapeutic Antibodies

Figure 1

Flow diagram of the crossmatch-ELISA for the detection of donor-specific anti-HLA class I antibodies. (a) Binding of the donor’s solubilized HLA class I molecules by monoclonal capture antibodies recognizing a monomorphic epitope on HLA class I molecules. (b) Binding of the donor-specific anti-HLA antibodies out of the recipient’s serum to the HLA molecules of the donor. (c) Binding of enzyme-conjugated secondary anti-human IgG (anti-human IgG/M/A) antibodies to the bound recipient’s donor-specific anti-HLA class I antibodies and subsequent color reaction. (d) Lysate control using an enzyme-conjugated monoclonal antibody directed against a second monomorphic epitope (AMS-ELISA) or the 2-microglobulin (AbCross) for detection in order to confirm the immobilization of a sufficient amount of HLA molecules by the capture antibody to generate a signal. The ELISA-variant for the detection of donor-specific anti-HLA class II antibodies is correspondingly designed.

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