The epitopic ''fingerprint'' of thyroid peroxidase-specific Fab isolated from a patient's thyroid gland by the combinatorial library approach resembles that of autoantibodies in the donor's serum

A new thyroid peroxidase (TPO)-specific Fab (KM1) was obtained from an immunoglobulin gene combinatorial library of patient KM containing L chain genes amplified with a single ''promiscuous'' V kappa oligonucleotide primer. The KM1 L chain is encoded by a mutated B3 gene (V kappa IV family). Another mutated B3 L chain had keen identified previously in a TPO-specific Fab (WR1.223) isolated from a different patient (WR). In contrast to patient KM, the WR L chains were amplified with a panel of V kappa family-specific primers. Both KM1 and WR1.223 bind TPO with high affinity (similar to 1 x 10(-9) M) and interact with an epitope in the B domain of the TPO immunodominant region. TPO-specific Fab previously isolated from a WR combinatorial library constructed with the promiscuous V kappa primer recognized the TPO A domain and none used a B3-like L chain. Remarkably, for both patients, Fab isolated from L chains generated with the promiscuous V kappa primer had epitopic profiles similar to autoantibodies in the donor's serum (RM-B domain; WR-A domain). Our data indicate that the promiscuous primer preferentially amplifies the dominant L chain present in vivo. However, to obtain a relatively rare Fab (such as the B domain Fab from WR), family-specific kappa primers are required. These findings provide insight into the relationship between TPO autoantibody gene usage, epitopic recognition, and the effectiveness of the combinatorial library approach. (C) 1997 Academic Press.