对超过中蛋白质选损失的缓冲效应及其与生物物理性质的关系
Aylin Mohammadzadehmarandi1, Andrew L Zydney1
1Department of Chemical Engineering, The Pennsylvania State University, University Park, Pennsylvania, USA.
Biotechnology progress
|May 23, 2024
概括
缓冲区选择显著影响超过过程中的蛋白质保留. 酸盐缓冲剂减少了牛血清白蛋白保留,与乙酸,酸和酸不同,这是由于影响水力动力直径的形状变化造成的.
科学领域:
- 生物物理化学 生物物理化学
- 蛋白质的配方 蛋白质的配方
- 分离科学 分离科学
背景情况:
- 蛋白质配方的有效超过/透过 (UF/DF) 要求具有高蛋白质保留力的膜.
- 缓冲区选择是一个关键的过程参数,可以影响 UF/DF 期间的蛋白质行为.
研究的目的:
- 研究不同缓冲器在超过过程中对牛血清白蛋白 (BSA) 保持的影响.
- 为了将缓冲器引起的蛋白质构造和水力动力直径的变化与UF/DF保留相关联.
主要方法:
- 使用模型蛋白 (BSA) 在各种缓冲剂 (乙酸盐,酸盐,酸胺,酸盐) 中的超过实验,pH值为4.8.
- 动态光散射 (DLS) 用于测量BSA水力动力直径的变化.
- 循环二元化 (CD) 光谱和差分扫描热量计 (DSC) 来评估蛋白质的结构稳定性.
主要成果:
- 与酸盐,酸盐和酸胺缓冲剂相比,BSA保留在4.8pH的酸盐缓冲剂中明显较低.
- DLS表明酸盐缓冲区BSA水力动力直径的小但显著下降.
- CD和DSC的结果表明,酸盐缓冲区内BSA的细微形状变化.
结论:
- 特定的缓冲组合可以影响蛋白质构成和水力动力学尺寸,从而影响在UF/DF过程中膜保留.
- 在pH值4.8的酸盐缓冲剂诱导了BSA的变化,导致保留减少,突出显示了缓冲剂选择在蛋白质加工中的重要性.
相关概念视频
Dialysis
641
Dialysis is a diffusion-based purification process that separates analyte molecules from a complex matrix. This is accomplished by allowing molecules in the solution to pass through a semipermeable membrane into a liquid on the other side. The membrane is usually made of cellulose acetate or cellulose nitrate, and the second liquid must be miscible with the solution. Ions (e.g., chloride or sodium) or organic molecules (e.g., glucose) can pass through the membrane pores, which generally have...
641
Factors Affecting Renal Clearance: Drug's Physicochemical Properties and Plasma Levels
211
Renal clearance of a drug is influenced by various factors, including its physicochemical properties and plasma levels. These factors play a significant role in determining how efficiently the kidneys eliminate a drug.
One important factor is the drug's molecular size. The kidneys readily excrete smaller molecules below 300 Daltons (Da). On the other hand, molecules weighing between 300 and 500 Da are excreted through both urine and bile. Larger molecules above 500 Da tend to be excreted...
One important factor is the drug's molecular size. The kidneys readily excrete smaller molecules below 300 Daltons (Da). On the other hand, molecules weighing between 300 and 500 Da are excreted through both urine and bile. Larger molecules above 500 Da tend to be excreted...
211
Size-Exclusion Chromatography
560
In size-exclusion chromatography (SEC), also known as molecular-exclusion or gel-permeation chromatography, molecules are separated based on their sizes. This technique is important for separating large molecules such as polymers and biomolecules. The two classes of micron-sized stationary phases encountered in SEC are silica particles and cross-linked polymer resin beads. Both materials are porous, but their pore sizes vary significantly.
Silica particles offer advantages such as rigidity,...
Silica particles offer advantages such as rigidity,...
560
Centrifugation
2.2K
Centrifugation is a separation technique based on differences in density or size. It is commonly used to separate solids from aqueous interferents. During centrifugation, the sample is placed in centrifugation tubes and spun at high angular velocity, which allows centrifugal force to act differentially on the different densities or masses of the components. After spinning, the supernatant liquid is decanted. Depending on the specific application, either the pellet or the supernatant is retained...
2.2K
Detergent Purification of Membrane Proteins
5.2K
Detergents are used to purify the integral proteins of the membrane. The hydrophobic portion of the detergent can replace membrane phospholipids while solubilizing the membrane proteins. When detergent monomers reach a specific concentration in a solution called critical micelle concentration (CMC), they form micelles. Above CMC, the concentration of the detergent monomers remains in equilibrium with the micelle. The number of detergent monomers present in the CMC varies for each detergent, and...
5.2K
Physiological Pharmacokinetic Models: Assumption with Protein Binding
41
Physiological models with protein binding in pharmacokinetics offer a sophisticated approach to understanding drug disposition. These models consider drug-protein interactions, enabling them to effectively predict drug concentrations in different organs and tissues. This precision aids in accurate drug dosing, providing a significant advantage over conventional models. A key process within these models is equilibration, which ensures that drug concentrations achieve a steady state within the...
41


