Neutralization and purification of thyroid stimulating antibody (TSAb) and thyroid blocking antibody (TBAb) by heterophilic antibody to animal IgG in Graves' disease

Immunological similarity of thyroid stimulating antibody (TSAb) and thyroid blocking antibody (TBAb) with animal IgG

“…The fact that Fab portion of LATS-IgG may have immunological similarity with bTSH (mammalian TSH) compared to hTSH was described in the previous paper [4]. These facts are also the supporting evidence for our findings that Fab of TSAb-and TBAb-IgG have immunological similarity with bFab compared to hFab.…”

“…Affinty chromatography of TSAb-and TBAb-IgG (purified by Protein A) on these anti-animal IgG Abs-bound column and also TSAband TBAb-Fab, -F(ab') 2 and -IgG on anti-bF(ab') 2 Ab-bound column was performed as described previously [4].…”

“…Rabbit anti-human (h) IgG antibody (made by our Lab.) showed precipitation line against both hIgG and monkey IgG, but negative against other mammalian IgGs, although cross-reactivity with these mammalian IgGs was observed [4]. Goat anti-bF(ab') 2 Ab was obtained from Rockland Inc. (Gilbertville, PA, USA).…”

“…Then, the supernatant fraction obtained by centrifugation after adding PEG was used for TBII and TSAb assay as described previously [4].…”

“…Lately, we found neutralization of TSAb and TBAb activity by heterophilic antibody to animal IgG (without antibody to human) in patients with Graves' disease and developed affinity purification of TSAb and TBAb-IgG by this anti-animal IgG antibody [4]. In the present experiment we examined immunological similarity of TSAb and TBAb with animal IgG using anti-animal IgG (dog, bovine, porcine, rabbit) Ab produced experimentally and immunological similarity of TSAb-and TBAb-Fab with bovine Fab using goat anti-bovine F(ab') 2 Ab.…”