Measures to overcome HAMA interferences in immunoassays.

The incidence of human anti-mouse antibodies (HAMA) in different patient sera panels may be debatable, the interference of HAMA, if present, in immunochemically based assays is, however, a fact. This interference can lead to falsely elevated or depressed results depending on the nature of the HAMAs involved and the particular assay format chosen. For several reasons, assays for serum tumor markers may be especially prone to HAMA interference. Consequently, in the development of such assays, special attention should be given to the HAMA issue. In our experience, the degree of HAMA interference varies greatly from one assay to another. Nevertheless, the HAMA issue has to be addressed. Several methods have been described to remove HAMA (and other interfering substances) via sample pretreatment. Alternatively, there are also some options to counterbalance the potential effect of HAMA by using assay reagents optimized for minimal HAMA susceptibility, e.g. inclusion of an excess of non-relevant mouse antibodies. However, there is no guarantee that any given assay will not be affected by HAMA. This is especially true if a portion of the HAMA in a patient's sample is comprised of anti-idiotypic "internal image" antibodies.