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pH值和盐对混合模式色谱中溶菌酶吸附的影响。

Influence of pH value and salts on the adsorption of lysozyme in mixed-mode chromatography.

作者信息

Kreusser Jannette, Jirasek Fabian, Hasse Hans

机构信息

Laboratory of Engineering Thermodynamics (LTD) TU Kaiserslautern Kaiserslautern Germany.

出版信息

Eng Life Sci. 2021 Nov 8;21(11):753-768. doi: 10.1002/elsc.202100058. eCollection 2021 Nov.

DOI:10.1002/elsc.202100058
PMID:34764827
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8576077/
Abstract

Mixed-mode chromatography (MMC) is an interesting technique for challenging protein separation processes which typically combines adsorption mechanisms of ion exchange (IEC) and hydrophobic interaction chromatography (HIC). Adsorption equilibria in MMC depend on multiple parameters but systematic studies on their influence are scarce. In the present work, the influence of the pH value and ionic strengths up to 3000 mM of four technically relevant salts (sodium chloride, sodium sulfate, ammonium chloride, and ammonium sulfate) on the lysozyme adsorption on the mixed-mode resin Toyopearl MX-Trp-650M was studied systematically at 25℃. Equilibrium adsorption isotherms at pH 5.0 and 6.0 were measured and compared to experimental data at pH 7.0 from previous work. For all pH values, an exponential decay of the lysozyme loading with increasing ionic strength was observed. The influence of the pH value was found to depend significantly on the ionic strength with the strongest influence at low ionic strengths where increasing pH values lead to decreasing lysozyme loadings. Furthermore, a mathematical model that describes the influence of salts and the pH value on the adsorption of lysozyme in MMC is presented. The model enables predicting adsorption isotherms of lysozyme on Toyopearl MX-Trp-650M for a broad range of technically relevant conditions.

摘要

混合模式色谱法（MMC）是一种用于挑战性蛋白质分离过程的有趣技术，它通常结合了离子交换（IEC）和疏水相互作用色谱法（HIC）的吸附机制。MMC中的吸附平衡取决于多个参数，但对其影响的系统研究却很少。在本工作中，系统研究了pH值和高达3000 mM的四种技术相关盐（氯化钠、硫酸钠、氯化铵和硫酸铵）的离子强度对溶菌酶在混合模式树脂Toyopearl MX-Trp-650M上吸附的影响，实验温度为25℃。测量了pH 5.0和6.0时的平衡吸附等温线，并与之前工作中pH 7.0时的实验数据进行了比较。对于所有pH值，均观察到随着离子强度增加，溶菌酶负载量呈指数下降。发现pH值的影响显著取决于离子强度，在低离子强度下影响最强，此时pH值升高会导致溶菌酶负载量降低。此外，还提出了一个描述盐和pH值对MMC中溶菌酶吸附影响的数学模型。该模型能够预测在广泛的技术相关条件下，溶菌酶在Toyopearl MX-Trp-650M上的吸附等温线。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/62862fe250f1/ELSC-21-753-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/f41d4468af4e/ELSC-21-753-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/7928c17272a4/ELSC-21-753-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/bbbca213ebb2/ELSC-21-753-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/3b4705ed4897/ELSC-21-753-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/00b4500f9a09/ELSC-21-753-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/6e1984e5aeb3/ELSC-21-753-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/62862fe250f1/ELSC-21-753-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/f41d4468af4e/ELSC-21-753-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/7928c17272a4/ELSC-21-753-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/bbbca213ebb2/ELSC-21-753-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/3b4705ed4897/ELSC-21-753-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/00b4500f9a09/ELSC-21-753-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/6e1984e5aeb3/ELSC-21-753-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43da/8576077/62862fe250f1/ELSC-21-753-g004.jpg

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J Chromatogr A. 2017 Oct 27;1521:73-79. doi: 10.1016/j.chroma.2017.09.024. Epub 2017 Sep 14.
3
Influence of mixed electrolytes on the adsorption of lysozyme, PEG, and PEGylated lysozyme on a hydrophobic interaction chromatography resin.
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A mixed-mode resin with tryptamine ligand for human serum albumin separation.一种用于人血清白蛋白分离的含色胺配体的混合模式树脂。
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