H2 domain name had been extremely at the mercy of cures, with those in this new V

Liu et al. (2010a) determined the relative susceptibility of all of the disulfide bonds of a human https://www.datingmentor.org/tr/mamba-inceleme IgG₁ to reduction, and found that inter-chain disulfide bridges between the heavy chain (HC) and the light chain (LC) were the most labile, followed by the two HC–HC disulfide bridges of the hinge (the most N-terminal of which was more susceptible than the other). _L, C_L, V_H, and C_H1 domains, and finally the C_H3 domain, which was most stable to reduction ( Liu et al., 2010a ). They also determined that the LC–HC inter-chain bonds of IgG₁? were more labile than those of IgG₁? ( Liu et al., 2010a ), a result similar to that found by Montano and Morrison (2002) with IgG₂? vs IgG₂?. The C-terminus of C? ends in the Cys residue that bridges to the HC, but with C_?, the disulfide bridging Cys residue is followed by a Ser residue, which is the C-terminal residue of that chain ( Liu et al., 2011a ). The presence of that Ser residue (or even if changed to an Ala residue) was found to be the cause for the increase in lability of the IgG₁? HC–LC disulfide bridge. Deletion of that C-terminal residue increased the stability of the IgG₁? HC–LC disulfide bridge ( Liu et al., 2011a ).

As noted in Section 9 , person IgG

₂ antibodies have the ability to form multiple structural isoforms due to alternative disulfide paring ( Wypych et al., 2008; ; Liu et al., 2008c; Allen et al., 2009; Zhang et al., 2010; Lightle et al., 2010 ) (see Figure 16.3 ). These isoforms are found not only in recombinant IgG₂ antibodies, but also in IgG₂s isolated from myeloma patients and normal human serum. Thus they are “natural” and an intrinsic property of the human IgG₂ isotype, rather than being the product of recombinant DNA manipulations and/or cell culture production ( Wypych et al., 2008 ). The root cause of this alternative disulfide bridging appears to be due mostly to the presence of two extra disulfide bonds in the hinge of IgG₂s ( ). The three major isomers have been labeled IgG₂-A (i.e. “normal” IgG₂ disulfide bridging), IgG₂-A/B, and IgG₂-B ( Wypych et al., 2008; Lightle et al., 2010 ; see Figure 16.3 ). Additional minor isoforms have also been described, including IgG₂-B₁ and IgG₂B₂ ( ), and, more recently IgG₂-B₃ ( Zhang et al., 2010 ). These isoforms of IgG₂ are generated because the Cys residue in the LC can form a disulfide bond with any of the three different Cys residues in the HC, Cys127 (Cys131; EU numbering) of the C_H1 domain or Cys232 (Cys219; EU numbering) or Cys233 (Cys220; EU numbering) in the hinge region ( Allen et al., 2009 ).

Multiple investigators have demonstrated that the major species of recombinant IgG₂ is not the “normal” IgG₂-A ( Wypych et al., 2008; c; Zhang et al., 2010 ). Liu et al. (2008b) demonstrated that B cells produce IgG₂-A, which is converted relatively rapidly in serum to IgG₂-A/B, followed by slower conversion to IgG₂-B, prompting the authors to state that the form might be an indicator of the “age” of an IgG₂ in serum. Liu et al. (2008c) also demonstrated that the clearance of the different forms of IgG₂ were essentially identical. The significance of this is that the different isoforms do not have the same binding activity of functionality. Dillon et al. (2008) compared an IgG₁ isotype with the wild-type IgG₂ and its various isoforms of an anti-IL-1? MAb. They found that IgG₁ had about a threefold greater activity than classical IgG₂ (IgG₂-A) and its various isoforms, and that the IgG₂-B isoform had about a threefold lower affinity and a threefold higher IC₅₀ than IgG₂-A, indicating that there are significant differences in the in vitro biological activity of the various isoforms ( Dillon et al., 2008 ). Allen et al. (2009) generated Cys > Ser mutations of several of the IgG₂ Cys residues involved in multiple isoform generation, and showed that the mutant IgG₂ molecules were both stable and biologically active.