Preparation, characterization and antigenicity of soybean 7S globulin-jujube polysaccharide complexes

doi:10.3969/j.issn.1000-1565.2024.03.004

Abstract

Abstract: In order to reduce the antigenicity of soybean, the soybean 7S globulin-Jujube polysaccharide(7S-JP)complexes were prepared by glycosylation reaction. The effects of reaction temperature, reaction time, and material ratio of 7S to JP on the structure of 7S-JP complexes were studied by asymmetrical flow field-flow fractionation coupled with online ultraviolet-visible detector, multi-angle light scattering detector, and differential refractive index detector(AF4-UV-MALS-dRI). Meanwhile, the radius of gyration, molar mass, and conformation of 7S-JP complexes were characterized by AF4-UV-MALS-dRI. The antigenicity of 7S-JP complexes was studied by indirect enzyme-linked immunosorbent assay. The results showed that: The 7S reacted with JP sufficiently at 70 ℃ in 20 min; The structure- DOI:10.3969/j.issn.1000-1565.2024.03.004大豆7S球蛋白-大枣多糖复合物的制备、表征及抗原性张晨婧¹,张西瑞²,郭子瑶¹,韩丹丹²,窦海洋¹(1.河北大学基础医学院,河北省炎性自身免疫性疾病发病机制及防治重点实验室,河北保定 071000;2.河北大学公共卫生学院,河北保定 071000)摘要:为了降低大豆的抗原性,通过糖基化反应制备了大豆7S球蛋白-大枣多糖(7S-JP)复合物.采用非对称场流分离技术在线联用紫外可见光检测器、多角度光散射检测器和示差折光检测器(AF4-UV-MALS-dRI)研究糖基化反应温度、时间和物料比对7S-JP复合物结构的影响,对7S-JP复合物的回转半径、摩尔质量和构象进行表征;采用间接竞争酶联免疫吸附法探究7S-JP复合物的抗原性.AF4-UV-MALS-dRI结果表明:在反应温度为70 ℃、反应时间为20 min时,7S与JP糖基化反应效果最佳;当7S与JP的质量比为1∶3时,形成的7S-JP复合物结构紧密,有利于破坏其空间抗原表位,抗原抑制率为(69.1±0.99)%.关键词:大豆7S球蛋白;大枣多糖;非对称场流分离中图分类号:O657 文献标志码:A 文章编号:1000-1565(2024)03-0254-07Preparation, characterization and antigenicity of soybean 7S globulin-jujube polysaccharide complexesZHANG Chenjing¹, ZHANG Xirui², GUO Ziyao¹, HAN Dandan², DOU Haiyang¹(1. Key Laboratory of Pathogenesis Mechanism and Control of Inflammatory-Autoimmune Disease of Hebei Province, School of Basic Medical Sciences, Hebei University, Baoding 071000, China;2. College of Public Health, Hebei University, Baoding 071000, China)Abstract: In order to reduce the antigenicity of soybean, the soybean 7S globulin-Jujube polysaccharide(7S-JP)complexes were prepared by glycosylation reaction. The effects of reaction temperature, reaction time, and material ratio of 7S to JP on the structure of 7S-JP complexes were studied by asymmetrical flow field-flow fractionation coupled with online ultraviolet-visible detector, multi-angle light scattering detector, and differential refractive index detector(AF4-UV-MALS-dRI). Meanwhile, the radius of gyration, molar mass, and conformation of 7S-JP complexes were characterized by AF4-UV-MALS-dRI. The antigenicity of 7S-JP complexes was studied by indirect enzyme-linked immunosorbent assay. The results showed that: The 7S reacted with JP sufficiently at 70 ℃ in 20 min; The structure- 收稿日期:2024-01-24;修回日期:2024-03-12 基金项目:河北省重点研发项目(22372505D);河北省大学生创新创业训练计划项目(S202310075055);河北大学实验室开放项目(sy202245) 第一作者:张晨婧(2000—),女,河北大学在读硕士研究生,主要从事场流分离研究. E-mail: zhangccc10@163.com 通信作者:窦海洋(1983—),男,河北大学副教授,博士,主要从事场流分离研究.E-mail: douhaiyang-1984@163.com韩丹丹(1981—),女,河北大学教授,博士,主要从事卫生检验研究. E-mail: hdd@hbu.edu.cn第3期张晨婧等:大豆7S球蛋白-大枣多糖复合物的制备、表征及抗原性of 7S-JP complexes obtained with a material ratio of 1∶3 was more compact, which is beneficial to destroy its spatial epitope, and the antigen inhibition rate of 7S-JP(1∶3)achieved to be(69.1±0.99)%.

Key words: soybean 7S globulin, jujube polysaccharide, asymmetrical flow field-flow fractionation

CLC Number:

O657

ZHANG Chenjing, ZHANG Xirui, GUO Ziyao, HAN Dandan, DOU Haiyang. Preparation, characterization and antigenicity of soybean 7S globulin-jujube polysaccharide complexes[J]. Journal of Hebei University(Natural Science Edition), 2024, 44(3): 254-260.

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