相关实验视频

Updated: Jun 23, 2025

Chemical Triphosphorylation of Oligonucleotides

Chemical Triphosphorylation of Oligonucleotides

Published on: June 2, 2022

合成酸寡核酸二聚体二维序列选择性综合性奇拉×逆相液态色谱

Feiyang Li¹, Cornelius Knappe¹, Niklas Carstensen¹

¹Institute of Pharmaceutical Sciences, Pharmaceutical (Bio-)Analysis, University of Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany.

Journal of chromatography. A

|June 16, 2024

概括

酸修饰增强了寡核酸的稳定性,但产生了复杂的二聚体. 这项研究评估了液体染色学方法,发现使用奇拉柱的二维染色学显著改善了这些复杂的寡核酸混合物的分离.

科学领域:

制药化学制药化学的相关文章
分析化学分析化学
生物技术是生物技术.

背景情况:

基于核酸的药物,如寡核酸 (ONs),越来越普遍.
酸 (PS) 修饰改善了ON稳定性和细胞吸收,但引入了立体异构复杂性.
非立体选择性引入PS链接产生了众多的二元立体体,使质量控制复杂化并影响生物活性.

研究的目的:

为了研究各种液态染色学 (LC) 方法的立体选择性,以分离类酸寡核酸 (PSO) 双立体体.
评估不同LC技术的有效性,包括离子配对逆相 (IP-RPLC) 和合色谱,用于PSO分析.
为了解决有关PSO二体样本的批量复制性的质量和监管问题.

主要方法:

评估了IP-RPLC,RPLC和LC的二聚体选择性,用于模型PSO和PSO-GalNAc结合物,并使用基于基尔多糖的柱子.
采用序列选择性综合二维染色学 (SC 2D-LC),在第一维中使用基于粉素的性静止相 (CSP).
在SC 2D-LC设置中使用乙酸乙烯作为第二维的离子对RPLC.

主要成果:

标准LC方法显示,复杂的PSO二聚体混合物的分离功率有限.
SC 2D-LC显著提高了分离功率,解决了更多的二聚体峰值.
通过结合性CSP和IP-RPLC实现了改善的二聚体选择性.

结论:

序列选择性综合二维色谱是一种强大的工具,用于解决复杂的PSO二体混合物.
开发的LC方法提供了改进的立体选择性,这对于PS改性寡核酸的质量控制和监管合规性至关重要.
这种方法有助于更好地表征PSO药物物质,确保一致的批量质量和可预测的药理特性.

关键词:

氨三 ((α-甲基基碳酸盐)奇拉尔静止阶段的静止阶段迪亚斯特里欧默分离的分离.酸酸的寡核酸.两维液体色谱是二维的液体色谱.

更多相关视频

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Published on: July 28, 2017

Synthesis of Wavelength-shifting DNA Hybridization Probes by Using Photostable Cyanine Dyes

Synthesis of Wavelength-shifting DNA Hybridization Probes by Using Photostable Cyanine Dyes

Published on: July 6, 2016

相关概念视频

Chirality at Nitrogen, Phosphorus, and Sulfur

Chirality at Nitrogen, Phosphorus, and Sulfur

Chirality is most prevalent in carbon-based tetrahedral compounds, but this important facet of molecular symmetry extends to sp3-hybridized nitrogen, phosphorus and sulfur centers, including trivalent molecules with lone pairs. Here, the lone pair behaves as a functional group in addition to the other three substituents to form an analogous tetrahedral center that can be chiral.
A consequence of chirality is the need for enantiomeric resolution. While this is theoretically possible for all...

High-Performance Liquid Chromatography: Introduction

High-Performance Liquid Chromatography: Introduction

High-performance liquid chromatography(HPLC), formerly referred to as High-pressure liquid chromatography, is a powerful technique used to separate, identify, and quantify components in complex mixtures. The term "high pressure" refers to using high pressure to push the liquid mobile phase through the tightly packed columns.
In HPLC, two phases play a critical role in the separation process:

¹H NMR Chemical Shift Equivalence: Enantiotopic and Diastereotopic Protons

¹H NMR Chemical Shift Equivalence: Enantiotopic and Diastereotopic Protons

Replacing each alpha-hydrogen in chloroethane by bromine (or a different functional group) yields a pair of enantiomers. Such protons are called prochiral or enantiotopic and are related by a mirror plane. Enantiotopic protons are chemically equivalent in an achiral environment. Because most proton NMR spectra are recorded using achiral solvents, enantiotopic hydrogens yield a single signal.
In chiral compounds such as 2-butanol, replacing the methylene hydrogens at C3 produces a pair of...

Affinity Chromatography

Affinity Chromatography

Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...

Prochirality

Prochirality

The concept of prochirality leads to the nomenclature of the individual faces of a molecule and plays a crucial role in the enantioselective reaction. It is a concept where two or more achiral molecules react to produce chiral products. A typical process is the reaction of an achiral ketone to generate a chiral alcohol. Here, the achiral reactant reacts with an achiral reducing agent, sodium borohydride, to generate an equimolar mixture of the chiral enantiomers of the product. For example, an...

Stereoisomerism

Stereoisomerism

Isomerism in Complexes
Isomers are different chemical species that have the same chemical formula.
Transition metal complexes often exist as geometric isomers, in which the same atoms are connected through the same types of bonds but with differences in their orientation in space. Coordination complexes with two different ligands in the cis and trans positions from a ligand of interest form isomers. For example, the octahedral [Co(NH3)4Cl2]+ ion has two isomers (Figure 1) In the cis...