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Homobifunctional Phenylselenoester Cross-linker For The Preparation Of Protein Conjugates And Its Application In Synthetic Carbohydrate-based Vaccine

Posted on:2018-07-17Degree:MasterType:Thesis
Country:ChinaCandidate:X G YinFull Text:PDF
GTID:2321330518975711Subject:Organic Chemistry
Abstract/Summary:PDF Full Text Request
Protein conjugation with small molecule is popular in the fields of synthetic vaccine,antibody-drug conjugate and immunoassay. Homobifuctional cross-linker, symmetically containing same reactive group on both ends, is one of most useful protocols for conjugation. After conjugation with small molecular, the intermediate react with the active site of protein. An ideal cross-linker with high conjugation efficiency must have suitable stability and reactivity.Adipic acid-based homobifuctional cross-linker has been widely used in protein conjugation especially in synthetic vaccine, because its non-immunogenic feature would not affect the immune response to the hapten targeted. Adipic acid N-hydroxysuccinimide(NHS) diester, a typical cross-linker widely used in synthetic vaccine, is too reactive that tends to hydrolysed rapidly,leading to a low level of incorporation in protein. Its counterpart that terminated by p-nitrophenyl (PNP) ester has improved stability, but it is not reactive enough to achieve high conjugation efficiency. Phenylselenoester demonstrates an improved balance between reactivity and stability.This dissertation includes the following: (1) Both stability and reactivity of phenylselenoester are systematically investigated. (2) Adpic acid disenoester linker is employed in the conjugation of proteins with a variety of small molecules, including tumor-associated carbohydrate antigens (Tn, STn), tumor associated-peptide antigen(MUC-5), fluorescent dye (NBD), and nicotine. (3) The protein conjugate STn-OVA is used as anti-tumor vaccine, and its immunological test showed that it is able to induce the immune system, resulting in antibody class-switching from IgM to IgG. These results shows adpic acid disenoester cross-linker can couple haptens to protein with high efficacy under mild conditions,and it will provide a practical tool for synthetic vaccines and protein-related drugs.
Keywords/Search Tags:adipic acid diselenoester, protein conjugation, synthetic vaccine, tumor carbohydrate antigens
PDF Full Text Request
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