Bioinformatics Institute, Agency for Science, Technology and Research (A*STAR), Singapore.
Bioinformatics Institute, Agency for Science, Technology and Research (A*STAR), Singapore; p53 Laboratory, Agency for Science, Technology and Research (A*STAR), Singapore.
J Allergy Clin Immunol. 2019 Aug;144(2):514-523.e5. doi: 10.1016/j.jaci.2019.03.028. Epub 2019 Apr 14.
Variable heavy chain (VH) family frameworks (FWRs) have been reported to affect antibody receptor and superantigen binding; however, such effects in IgE remain largely unknown. Given that VH family biases have been previously reported in IgE of certain allergies, there is a need to investigate this phenomenon for biotechnological and therapeutic purposes.
We sought to investigate the effects of VH families on IgE interaction with FcεRIα, anti-IgE omalizumab, antigen, and superantigen protein A (spA) by using the pertuzumab and trastuzumab IgE models.
Pertuzumab VH1-VH7 family variants of IgE with the same complementarity-determining regions were investigated with regard to their binding interactions to FcεRIα, Her2, omalizumab, and spA. Notable FcεRIα-IgE observations were cross-checked against appropriate trastuzumab IgE VH variants. Computational structural modeling and simulations were also performed for insight into the mechanism of interactions with various VH FWRs.
The pertuzumab VH5 IgE variant, but not the trastuzumab VH5 IgE, was found to interact with FcεRIα significantly longer than the respective VH family variants within each model antibody. No significant differences in interaction were found between IgE and omalizumab for the pertuzumab VH variants. Although trastuzumab VH3 interacted with spA, none of our pertuzumab VH variants, including VH3, associated with spA.
We found unexpected varying allosteric communications caused by the VH family FWRs to the FcεRIα-, Her2-, and spA-binding regions of pertuzumab IgE, with implications for use of IgE/anti-IgE therapeutics to treat allergy and IgE therapeutics in allergo-oncology.
可变重链（VH）家族框架（FWR）已被报道会影响抗体受体和超抗原结合；然而，IgE 中的这种影响在很大程度上仍然未知。鉴于某些过敏症的 IgE 中先前已经报道了 VH 家族偏倚，因此有必要对此现象进行研究，以满足生物技术和治疗目的的需要。
我们试图通过使用曲妥珠单抗和 pertuzumab 的 IgE 模型，研究 VH 家族对 IgE 与 FcεRIα、抗 IgE 奥马珠单抗、抗原和超抗原蛋白 A（spA）相互作用的影响。
研究了具有相同互补决定区的 pertuzumab VH1-VH7 家族变体的 IgE 与其与 FcεRIα、Her2、奥马珠单抗和 spA 的结合相互作用。对 FcεRIα-IgE 的重要观察结果与适当的 trastuzumab IgE VH 变体进行了交叉检查。还进行了计算结构建模和模拟，以深入了解与各种 VH FWR 相互作用的机制。
与各自模型抗体中的其他 VH 家族变体相比，发现 pertuzumab VH5 IgE 变体而非 trastuzumab VH5 IgE 变体与 FcεRIα 的相互作用时间明显更长。在 pertuzumab VH 变体中，IgE 与奥马珠单抗之间的相互作用没有发现显著差异。虽然 trastuzumab VH3 与 spA 相互作用，但我们的 pertuzumab VH 变体中没有一个与 spA 相关，包括 VH3。
我们发现了出乎意料的由 VH 家族 FWR 引起的变构通讯，这些变构通讯会影响 pertuzumab IgE 的 FcεRIα、Her2 和 spA 结合区域，这对使用 IgE/抗 IgE 疗法治疗过敏症和 IgE 疗法在过敏肿瘤学中的应用具有影响。
