A new UltraScan module for the characterization and quantification of analytical buoyant density equilibrium experiments to determine AAV capsid loading

IF 2.2 4区 生物学 Q3 BIOPHYSICS European Biophysics Journal Pub Date : 2023-04-04 DOI:10.1007/s00249-023-01641-4
Alexey Savelyev, Emre H. Brookes, Amy Henrickson, Borries Demeler
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引用次数: 5

Abstract

A method for characterizing and quantifying peaks formed in an analytical buoyant density equilibrium (ABDE) experiment is presented. An algorithm is derived to calculate the concentration of the density forming gradient material at every point in the cell, provided the rotor speed, temperature, meniscus position, bottom of the cell position, and the loading concentration, molar mass, and partial specific volume of the density gradient-forming material are known. In addition, a new peak fitting algorithm has been developed which allows the user to automatically quantify the peaks formed in terms of density, apparent partial specific volume, and relative abundance. The method is suitable for both ionic and non-ionic density forming materials and can be used with data generated from the UV optical system as well as the AVIV fluorescence optical system. These methods have been programmed in a new UltraScan-III module (us_abde). Examples are shown that demonstrate the application of the new module to adeno-associated viral vector preparations and proteins.

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一个新的UltraScan模块,用于分析浮力密度平衡实验的表征和定量,以确定AAV衣壳负载
提出了一种分析浮力密度平衡(ABDE)实验中形成的峰的表征和定量方法。在已知转子转速、温度、半月板位置、槽底位置以及密度梯度成形材料的加载浓度、摩尔质量和部分比容的情况下,导出了一种计算密度梯度成形材料在槽内各点浓度的算法。此外,还开发了一种新的峰拟合算法,允许用户根据密度、表观部分比体积和相对丰度自动量化形成的峰。该方法既适用于离子密度形成材料,也适用于非离子密度形成材料,并可用于UV光学系统和AVIV荧光光学系统生成的数据。这些方法已在新的UltraScan-III模块(us_abde)中编程。举例说明了新模块在腺相关病毒载体制备和蛋白质中的应用。
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来源期刊
European Biophysics Journal
European Biophysics Journal 生物-生物物理
CiteScore
4.30
自引率
0.00%
发文量
43
审稿时长
6-12 weeks
期刊介绍: The journal publishes papers in the field of biophysics, which is defined as the study of biological phenomena by using physical methods and concepts. Original papers, reviews and Biophysics letters are published. The primary goal of this journal is to advance the understanding of biological structure and function by application of the principles of physical science, and by presenting the work in a biophysical context. Papers employing a distinctively biophysical approach at all levels of biological organisation will be considered, as will both experimental and theoretical studies. The criteria for acceptance are scientific content, originality and relevance to biological systems of current interest and importance. Principal areas of interest include: - Structure and dynamics of biological macromolecules - Membrane biophysics and ion channels - Cell biophysics and organisation - Macromolecular assemblies - Biophysical methods and instrumentation - Advanced microscopics - System dynamics.
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