Department of Protein Structure, Function and Design, Graduate School of Pharmaceutical Sciences, Kyushu University, Japan.
Faculty of Pharmaceutical Sciences, Sojo University, Japan.
Biochem Biophys Res Commun. 2021 Jun 18;558:114-119. doi: 10.1016/j.bbrc.2021.04.070. Epub 2021 Apr 26.
The CH domain is a critical element of the human Immunoglobulin G (IgG) constant region. Although the CH domain is the least stable domain in IgG, it is also a promising scaffold candidate for developing novel therapeutic approaches. Recently, we succeeded in preparing glycosylated and non-glycosylated CH domain in the host organism Pichia pastoris. Herein, we verified that glycosylation of the CH domain decreased both, its tendency to aggregate and its immunogenicity in mice, suggesting that aggregation and immunogenicity are related. In addition, we have produced in P. pastoris a stabilized version of the CH domain with and without glycan, and their propensity to aggregate evaluated. We found that stabilization alone significantly decreased the aggregation of the CH domain. Moreover, the combination of glycosylation and stabilization completely suppressed its aggregation behavior. Since protein aggregation is related to immunogenicity, the combination of glycosylation and stabilization to eliminate the aggregation behavior of a protein could be a fruitful strategy to generate promising immunoglobulin scaffolds.
CH 结构域是人类免疫球蛋白 G（IgG）恒定区的关键元件。尽管 CH 结构域是 IgG 中最不稳定的结构域，但它也是开发新型治疗方法的有前途的支架候选物。最近，我们成功地在毕赤酵母宿主中制备了糖基化和非糖基化的 CH 结构域。在此，我们验证了 CH 结构域的糖基化降低了其在小鼠中的聚集倾向和免疫原性，表明聚集和免疫原性是相关的。此外，我们在毕赤酵母中生产了具有和不具有聚糖的 CH 结构域的稳定版本，并评估了它们的聚集倾向。我们发现，仅稳定化就显著降低了 CH 结构域的聚集。此外，糖基化和稳定化的组合完全抑制了其聚集行为。由于蛋白质聚集与免疫原性有关，因此糖基化和稳定化的组合消除蛋白质的聚集行为可能是产生有前途的免疫球蛋白支架的有效策略。
