Thekey differencebetween affinity and ion exchange chromatography is thatwe can use affinity chromatography to separate charged or uncharged components in a mixture whereas we can use ion exchange chromatography to separate charged components in a mixture.
色谱法是一种我们可以用来分离混合物中所需组件的技术。有不同类型的类型liquid chromatography, gas chromatography等等。亲和色谱和离子交换色谱法是液相色谱的两个子类别。同样,在这些技术中,有两个阶段。即,他们是固定阶段和流动阶段. The purpose of these techniques is to separate the components, depending on the binding of the components, in the mobile phase on to the surface of the stationary phase.
内容
1.Overview and Key Difference
2.什么是亲和力色谱法
3.What is Ion Exchange Chromatography
4.并排比较 - 表格形式的亲和力与离子交换色谱法
5。概括
什么是亲和力色谱法?
亲和力色谱法是一种生化技术,我们用它根据这些成分之间的相互作用来分离混合物中的组件。
在这种情况下,我们使用的交互包括以下内容:
- 抗原抗体相互作用
- 酶 - 基底相互作用
- Receptor-ligand interactions
- Protein-nucleic acid interactions
在此技术中,我们将分子的分子特性用于该分离技术。在这里,我们允许所需的化合物通过氢键,离子相互作用,二硫键,疏水相互作用等。不与固定相相互作用的分子将首先洗脱。因此,我们可以将其与混合物分开。所需的化合物将保持固定相。因此,我们可以使用洗脱溶剂将其分离,并使其也将其分开。
Affinity chromatography is useful in purification and concentrating a substance from a mixture using a buffer solution. Also, it is helpful in reducing the unwanted substances in a mixture. When considering the apparatus that we use for this process, we should use a column filled with our stationary phase. Then, we should load the mobile phase which contains the biomolecules that we are going to separate. Next, allow them to bind with the stationary phase. Thereafter, using a wash buffer, we can separate the non-target biomolecules, but the target molecules should have a high affinity for the stationary phase in order to success the separation process.
What is Ion Exchange Chromatography?
Ion chromatography is a form of liquid chromatography in which we can analyze ionic substances. Often, we use it to analyze inorganic anions and cations (i.e. chloride and nitrate anions and potassium, sodium cations). Though it is less common, we can analyze organic ions as well. Moreover, we can use this technique for the purification of proteins because proteins are charged molecules at certain pH values. Here, we use a solid stationary phase to which the charged particles can attach. For example, we can use the resin polystyrene-divinylbenzene copolymers as the solid support.
为了进一步解释,固定相具有固定的离子,例如硫酸盐阴离子或第四纪胺阳离子。如果我们要保持该系统的中立性,则每种都应与对立面(相反电荷的离子)相关联。在此,如果柜台是阳离子,则我们将系统命名为阳离子交换树脂。但是,如果柜台是阴离子,则系统是阴离子交换树脂。
There are five major phases in an ion exchange process;
- Initial stage
- 目标的吸附
- Starting of elution
- 洗脱的尽头
- 再生
亲和力和离子交换色谱法有什么区别?
Affinity chromatography is a biochemical technique that we use to separate components in a mixture depending on the interactions between these components whereas ion chromatography is a form of liquid chromatography in which we can analyze ionic substances. Therefore, the major difference between affinity and ion exchange chromatography is that we can use ion exchange chromatography only for the separation of ionic substances while the affinity chromatography is capable of separating both charged and uncharged particles. When considering the working principle, the difference between affinity and ion exchange chromatography is that the affinity chromatography proceeds due to the fact that target molecules have a high affinity for the stationary phase. However, for ion exchange chromatography, target molecules have an opposite charge to that of the stationary phase surface.
以下信息图显示了亲和力和离子交换色谱法之间的差异作为并排比较。
概括– Affinity vs Ion Exchange Chromatography
In summary, affinity and ion exchange chromatography are two forms of liquid chromatographic techniques. The key difference between affinity and ion exchange chromatography is that we can use affinity chromatography to separate charged or uncharged components in a mixture whereas we can use ion exchange chromatography to separate charged components in a mixture.
Reference:
1. “Affinity Chromatography.” Wikipedia, Wikimedia Foundation, 5 Oct. 2018.Available here
2. Libretexts. “Ion-Exchange Chromatography.” Chemistry LibreTexts, Libretexts, 29 Dec. 2016.Available here
Image Courtesy:
1. PDCOOK的“镍树脂” - 自己的工作,(CC BY-SA 3.0)viaCommons Wikimedia
2.”Iex”By Daliak (Public Domain) viaCommons Wikimedia
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