Determination of degradation for sulfo-SIAB, SM(PEG)2, and sulfo-GMBS by RPLC-UV analysis

IF 4.3 3区材料科学 Q1 ENGINEERING, ELECTRICAL & ELECTRONIC

ACS Applied Electronic Materials Pub Date : 2024-08-30 DOI:10.1016/j.jpba.2024.116455

Melissa Resto, Alison Vinitsky, Nicole A. Schneck, Jeremy J. Wolff, Asif Shajahan, Nicole Cibelli, Yaqiu Zhang, Yile Li, Krishna Gulla, Daniel B. Gowetski, Jason G. Gall, Q. Paula Lei

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引用次数: 0

Abstract

Bi-functional N-Hydroxysuccinimide (NHS) linkers are widely used in the conjugation processes linking an immunogen with a carrier protein capable of boosting immunity. A potential vaccine candidate against HIV-1, called fusion peptide (FP), is covalently linked to the recombinant tetanus toxoid heavy-chain fragment C (rTTHC) via this type of linker. A reversed-phase liquid chromatography (RPLC-UV) method was used to monitor the linker’s degradation kinetics in various buffers, mimicking the steps in the conjugation process. The kinetics of the reactivities of the linkers are revealed in this study and can provide a good guidance to help effective conjugation process before these linkers are completely hydrolyze to the inactive degradants. Three cross-linkers degradation pathways were evaluated: Sulfosuccinimidyl (4-iodoacetyl) aminobenzoate (Sulfo-SIAB), PEGylated SMCC (SM(PEG)₂), and N-γ-maleimidobutyryl-oxysulfosuccinimide ester (Sulfo-GMBS). We have reported kinetics for Sulfo-SIAB.

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通过 RPLC-UV 分析确定磺基-SIAB、SM(PEG)2 和磺基-GMBS 的降解情况

双功能 N-羟基琥珀酰亚胺（NHS）连接体被广泛应用于连接免疫原与能够增强免疫力的载体蛋白的共轭过程中。一种名为融合肽（FP）的潜在 HIV-1 候选疫苗就是通过这种连接体与重组破伤风类毒素重链片段 C（rTTHC）共价连接的。采用反相液相色谱法（RPLC-UV）监测连接体在各种缓冲液中的降解动力学，模拟连接过程中的各个步骤。本研究揭示了连接体的反应动力学，可在这些连接体完全水解为无活性降解剂之前为有效的共轭过程提供良好的指导。本研究评估了三种交联剂的降解途径：硫代琥珀酰亚胺基（4-碘乙酰基）氨基苯甲酸酯（Sulfo-SIAB）、聚乙二醇化 SMCC（SM(PEG)2）和 N-γ-马来酰亚胺基丁酰氧基硫代琥珀酰亚胺酯（Sulfo-GMBS）。我们已经报告了 Sulfo-SIAB 的动力学。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

ACS Applied Electronic Materials Multiple-

CiteScore

7.20

自引率

4.30%

发文量

567