Aasim Muhammad, Hidayatullah Khan Muhammad, Bibi Noor Shad, Zaman Khan Nadir
Department of Biotechnology University of Malakand Chakdara Dir (Lower) Pakistan.
School of Life Sciences University of Science and Technology of China Hefei Anhui P. R. China.
Eng Life Sci. 2017 Nov 29;18(3):179-186. doi: 10.1002/elsc.201700097. eCollection 2018 Mar.
This part of work was done to explore the basic understanding of the adsorption chromatography by determining the interaction of selected model proteins ( = 5) to monolithic chromatographic materials, with varying densities of butyl and phenyl ligands. Surface energetics approach was applied to study the interaction behavior. The physicochemical properties of the proteins and monolithic chromatographic materials were explored by contact angle and zeta potential values. These values were used to study protein to monolith interaction under various operating conditions. Surface energetics approach allowed the calculation of interaction energy as a function of distance, i.e. energy minimum values. Calculations were performed at various conditions to analyze the effect of major operating parameters on the interaction strength. The interaction strength exposed the hydrophobic nature of the monoliths which increases with increasing ligand density. Further, interaction energy of proteins were higher with monolith with butyl ligand compared to monolith with phenyl ligand. For instance, lactoferrin interaction to monoliths with butyl represents more interaction, i.e. 24.38 kT as compared to monoliths with phenyl i.e. 23.28 kT, keeping lambda as 0.2 nm and salt concentration as 100 mM of ammonium sulphate. Hence, more energy and time will be consumed for elution of proteins immobilized to monoliths with butyl. Similarly, the effect of solid surface for proteins immobilization, effect of ligand density and effect of lambda showed some interesting insights on the interaction behavior. The knowledge generated from the present work will help in the basic understanding as well as development of an efficient, low cost downstream processing design and may mimic the real chromatographic experiments.
本部分工作旨在通过测定选定的模型蛋白（n = 5）与具有不同丁基和苯基配体密度的整体色谱材料之间的相互作用，来探索对吸附色谱的基本理解。采用表面能方法研究相互作用行为。通过接触角和zeta电位值探索了蛋白质和整体色谱材料的物理化学性质。这些值用于研究在各种操作条件下蛋白质与整体柱的相互作用。表面能方法允许计算作为距离函数的相互作用能，即能量最小值。在各种条件下进行计算，以分析主要操作参数对相互作用强度的影响。相互作用强度揭示了整体柱的疏水性质，其随配体密度的增加而增加。此外，与具有苯基配体的整体柱相比，蛋白质与具有丁基配体的整体柱的相互作用能更高。例如，乳铁蛋白与具有丁基的整体柱的相互作用更强，即24.38 kT，而与具有苯基的整体柱相比为23.28 kT，保持λ为0.2 nm，盐浓度为100 mM硫酸铵。因此，洗脱固定在具有丁基的整体柱上的蛋白质将消耗更多的能量和时间。同样，固体表面对蛋白质固定的影响、配体密度的影响和λ的影响对相互作用行为显示出一些有趣的见解。从本工作中获得的知识将有助于对高效、低成本下游加工设计的基本理解和开发，并可能模拟实际的色谱实验。
