A systematic mutational analysis identifies a 5‐residue proline tag that enhances the in vivo immunogenicity of a non‐immunogenic model protein,FEBS Open Bio

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A systematic mutational analysis identifies a 5‐residue proline tag that enhances the in vivo immunogenicity of a non‐immunogenic model protein
FEBS Open Bio ( IF 2.8 ) Pub Date : 2020-07-29 , DOI: 10.1002/2211-5463.12941
Nafsoon Rahman ₁ , Mohammad Monirul Islam ₂ , Md Golam Kibria ₁ , Satoru Unzai ₃ , Yutaka Kuroda ₁

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Poor immunogenicity of small proteins is a major hurdle in developing vaccines or producing antibodies for biopharmaceutical usage. Here, we systematically analyzed the effects of 10 solubility controlling peptide tags (SCP‐tags) on the immunogenicity of a non‐immunogenic model protein, bovine pancreatic trypsin inhibitor (BPTI‐19A; 6 kDa). CD, fluorescence, DLS, SLS, and AUC measurements indicated that the SCP‐tags did not change the secondary structure content nor the tertiary structures of the protein nor its monomeric state. ELISA results indicated that the 5‐proline (C5P) and 5‐arginine (C5R) tags unexpectedly increased the IgG level of BPTI‐19A by 240‐ and 73‐fold, respectively, suggesting that non‐oligomerizing SCP‐tags may provide a novel method for increasing the immunogenicity of a protein in a highly specific manner.

中文翻译：

系统突变分析确定了一个 5 残基脯氨酸标签，可增强非免疫原性模型蛋白的体内免疫原性

小蛋白的免疫原性差是开发疫苗或生产用于生物制药的抗体的主要障碍。在这里，我们系统地分析了 10 种溶解度控制肽标签 (SCP-tags) 对非免疫原性模型蛋白牛胰蛋白酶抑制剂 (BPTI-19A; 6 kDa) 免疫原性的影响。CD、荧光、DLS、SLS 和 AUC 测量表明 SCP 标签没有改变蛋白质的二级结构含量、三级结构及其单体状态。ELISA 结果表明，5-脯氨酸 (C5P) 和 5-精氨酸 (C5R) 标签出人意料地将 BPTI-19A 的 IgG 水平分别提高了 240 倍和 73 倍，这表明非寡聚 SCP-标签可能提供一种新的以高度特异性的方式增加蛋白质的免疫原性的方法。

更新日期：2020-10-02

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