What is the Difference Between Gel Filtration and Affinity Chromatography

Thekey difference between gel filtration and affinity chromatographyis that gel filtration chromatography depends on the differences in molecular weight or size of the analyte sample, whereas affinity chromatography depends on the affinity of an analyte to an immobilized ligand.

The term chromatography in analytical chemistry refers to the process of the separation of components in a mixture using different techniques. Chromatography is a collective name that represents a various number of different methods for separation.

内容

1.Overview and Key Difference
2.What is Gel Filtration Chromatography
3.What is Affinity Chromatography
4.Gel Filtration vs Affinity Chromatography in Tabular Form
5。摘要 - 凝胶过滤与亲和力色谱法

什么是凝胶过滤色谱法？

凝胶过滤色谱法是一个分析侦探hnique in which the separation of components depends on the difference in molecular weight or size. This technique is also known as size-exclusion chromatography or molecular-sieve chromatography. The separation technique in this process depends on the differing ability of molecules in the sample to enter into the pores of the gel-filtration medium.

在凝胶过滤技术中固定阶段contains beads of hydrated, sponge-like material having pores of molecular dimensions containing a narrow range of sizes. If we pass an aqueous solution consisting of molecules of various sizes through a column containing these “molecular sieves,” the molecules that are larger than the pores of the filtration medium move quickly through the column. In contrast, the smaller molecules enter the pores of the gel and tend to move slowly through the column. Molecules elute from the column in order of decreasing molecular size. Here, the exclusion limit of the gel is the molecular mass of the smallest molecules unable to penetrate the pores of a given gel.

There are different types of gel filtration techniques, such as indirect gel filtration method, steady-state gel filtration, gel bead dialysis, etc. Indirect gel filtration chromatography is useful in the determination of free thyroid hormones. Steady-state gel filtration chromatography is an indirect technique useful mostly for the measurement of free steroids like cortisol, testosterone, etc. Gel bead dialysis, on the other hand, is a modification of steady-state gel filtration that is comparatively simpler.

What is Affinity Chromatography?

亲和力色谱法是一种分析技术和一种分离方法，取决于固定的特定结合相互作用ligandand its binding partner. Some examples include antibody-antigen binding, enzyme-substrate binding, and enzyme-inhibitor binding. Therefore, this technique has many important applications in nucleic acid purification, protein purification from cell extracts, and purification from blood.

在此技术中，最重要的特性是配体固定。我们可以为此使用各种材料，例如丙烯酸酯和硅胶。重要的是防止靶分子对配体的空间干扰。此外，抑制剂附着在固相上。我们将此抑制剂称为垫片。从经典上讲，垫片由烃链组成。

亲和力色谱的固定相包含核心，间隔和配体。有时，它还具有与配体耦合的金属离子。该技术中固定相的最优选的固相是琼脂糖凝胶，因为它很容易分配以填充并用任何大小的树脂床包装柱，并且足够大，可以使生物分子自由地流入珠子和珠子。配体可以以各种方式共价附着在珠聚合物上。最常见的垫片化合物是氰基溴，环氧化物，带有C6酸的环氧和二氧化物氧化物。另一方面，我们可以根据靶靶标有不同的配体，例如抗体 - 抗原，铁或铝离子 - 磷蛋白，抗原蛋白 - 生物素，谷胱甘肽-GST，螯合剂-HIS-HIS-HIS-HIS-HIS-HIS-TAGGER蛋白等。

What is the Difference Between Gel Filtration and Affinity Chromatography?

Chromatography is an important analytical technique. Gel filtration chromatography and affinity chromatography are two important variations of chromatography. The key difference between gel filtration and affinity chromatography is that gel filtration chromatography depends on the differences in molecular weight or size of the analyte sample, whereas affinity chromatography depends on the affinity of an analyte to an immobilized ligand.

摘要 - 凝胶过滤与亲和力色谱法

There are different types of chromatographic techniques according to their application and the nature of the analyte sample. Gel filtration and affinity chromatography are two such types of techniques. The key difference between gel filtration and affinity chromatography is that gel filtration chromatography depends on the differences in molecular weight or size of the analyte sample, whereas affinity chromatography depends on the affinity of an analyte to an immobilized ligand.