Anti-anti-idiotypic (Ab3) antibodies that bind progesterone-11alpha-bovine serum albumin differ in their combining sites from antibodies raised directly against the antigen.

Anti-anti-idiotypic (Ab3) antibodies that bind progesterone-11alpha-bovine serum albumin differ in their combining sites from antibodies raised directly against the antigen. post thumbnail image
Polyclonal rabbit anti-idiotypic (Ab2) antibodies raised in opposition to the antiprogesterone mAb DB3 (Ab1) had been used to induce an Ab3 antiprogesterone response in BALB/c mice. Whereas the affinity of Ab3 sera for progesterone was 10-50-times decrease than that of DB3, their steroid-binding specificity confirmed appreciable similarity to DB3. Two immunoglobulin M (IgM) Ab3 monoclonal antibodies (mAbs), 1A4 and 3B11, had been obtained, each of which sure progesterone conjugated to bovine serum albumin (progesterone-BSA).
1A4 additionally sure free progesterone, though with low affinity and really broad cross-reactivity. Like DB3, 1A4 is encoded by a heavy-chain variable area (VH) gene phase from the small VGAM3.Eight household, a restriction that’s attribute of antibodies raised in opposition to progesterone-11alpha-BSA. In distinction, 3B11 binds progesterone-11alpha-BSA however not free progesterone and is encoded by an unrelated VH gene from the J558 household.
The sunshine chain variable area (VL) of 1A4 lacks the intradomain disulphide bridge owing to substitute of CysL23 by Tyr. Each the 1A4 and 3B11 heavy chains have extraordinarily brief complementarity figuring out area (CDR) H3 loops, comprising three and 4 amino acids, respectively. Modelling of the combining website of 1A4 from the X-ray crystallographic construction of DB3 signifies that the brief H3 loop is a significant component within the lack of affinity and specificity for steroid.

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