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SARS-CoV-2 Peptide Bioconjugates Designed for Antibody Diagnostics
Bioconjugate Chemistry ( IF 4.7 ) Pub Date : 2021-06-28 , DOI: 10.1021/acs.bioconjchem.1c00186
Ivan M Ryzhov 1 , Alexander B Tuzikov 1 , Alexey V Nizovtsev 1 , Ludmila K Baidakova 2 , Oxana E Galanina 1 , Nadezhda V Shilova 1, 3 , Marina M Ziganshina 3 , Nataliya V Dolgushina 3 , Guldana R Bayramova 3 , Gennady T Sukhikh 3 , Eleanor C Williams 4 , Radhika Nagappan 4 , Stephen M Henry 4 , Nicolai V Bovin 1, 4
Affiliation  

In the near future, the increase in the number of required tests for COVID-19 antibodies is expected to be many hundreds of millions. Obviously, this will be done using a variety of analytical methods and using different antigens, including peptides. In this work, we compare three method variations for detecting specific immunoglobulins directed against peptides of approximately 15-aa of the SARS-CoV-2 spike protein. These linear peptide epitopes were selected using antigenicity algorithms, and were synthesized with an additional terminal cysteine residue for their bioconjugation. In two of the methods, constructs were prepared where the peptide (F, function) is attached to a negatively charged hydrophilic spacer (S) linked to a dioleoylphosphatidyl ethanolamine residue (L, lipid) to create a function–spacer–lipid construct (FSL). These FSLs were easily and controllably incorporated into erythrocytes for serologic testing or in a lipid bilayer deposited on a polystyrene microplate for use in an enzyme immunoassays (EIA). The third method, also an EIA, used polyacrylamide conjugated peptides (peptide-PAA) prepared by controlled condensation of the cysteine residue of the peptide with the maleimide-derived PAA polymer which were immobilized on polystyrene microplates by physisorption of the polymer. In this work, we describe the synthesis of the PAA and FSL peptide bioconjugates, design of test systems, and comparison of the bioassays results, and discuss potential reasons for higher performance of the FSL conjugates, particularly in the erythrocyte-based serologic assay.

中文翻译:

专为抗体诊断而设计的 SARS-CoV-2 肽生物偶联物

在不久的将来,COVID-19 抗体所需的测试数量预计将增加数亿。显然,这将使用各种分析方法和不同的抗原(包括肽)来完成。在这项工作中,我们比较了用于检测针对 SARS-CoV-2 刺突蛋白的大约 15-aa 的肽的特定免疫球蛋白的三种方法变化。这些线性肽表位是使用抗原性算法选择的,并用额外的末端半胱氨酸残基合成以进行生物缀合。在其中两种方法中,制备的构建体是将肽(F,功能)连接到带负电荷的亲水性间隔物(S),该​​间隔物与二油酰磷脂酰乙醇胺残基(L,脂质)相连,以创建功能-间隔物-脂质构建物(FSL )。这些 FSL 很容易且可控地掺入红细胞中以进行血清学测试或沉积在聚苯乙烯微板上的脂质双层中以用于酶免疫分析 (EIA)。第三种方法,也是 EIA,使用聚丙烯酰胺偶联肽(肽-PAA),通过肽的半胱氨酸残基与马来酰亚胺衍生的 PAA 聚合物的受控缩合制备,通过聚合物的物理吸附将其固定在聚苯乙烯微板上。在这项工作中,我们描述了 PAA 和 FSL 肽生物偶联物的合成、测试系统的设计和生物测定结果的比较,并讨论了 FSL 偶联物性能更高的潜在原因,特别是在基于红细胞的血清学测定中。
更新日期:2021-08-19
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