In general, the term immobilization refers to the act of the limiting movement or making incapable of movement. Immobilization is a universal phenomenon existing in nature. In the fields of biological and biomedical, many molecules such as biopolymers and small molecules are artificially immobilized on solid support as important research tools. The solid support should exhibit good chemical and physical stability and contain functional groups that can be bound to the active molecule. Common solid supports are magnetic particles, agarose beads, nanoparticles, films, microplates, array surfaces, and strips. The main methods for immobilization are adsorption, covalent binding, entrapment, encapsulation, and crosslinking, and the appropriate method should be selected according to the research purpose. The formation of covalent bonds between the ligand and the support usually provides the stronger linkages compared with other immobilization methods. The types of functionalities used for ligand attachment are easily reactive components such as amines, sulfhydryls, aldehydes, and carboxylic acids.
Amerigo Scientific offers SepSphere™ kits for the immobilization of small molecules and large biomolecules (including proteins, antibodies, peptides, and oligonucleotides) on agarose beads. All processes of immobilization, washing of unreacted starting materials, and column packing are performed in a column. The required consumables and reagents are available in the SepSphere™ kit. The washing is performed using a simple manual push syringe without the need for gas/air flow or vacuum.
Product | Ligand | Ligand Reactive Groups | Agarose Reactive Groups | Linkage | Stability of Linkage |
---|---|---|---|---|---|
SepSphere™ Antibody/Protein/Enzyme Immobilization Kit via Glyoxyl | Antibody/Protein/Enzyme | -NH2 | -C(=O)H (glyoxyl) | Secondary amine | Stable |
SepSphere™ Antibody/Protein/Enzyme Immobilization Kit via Glutaraldehyde | Antibody/Protein/Enzyme | -NH2 | Glutaraldehyde | Imine | Stable at pH 5-9 |
SepSphere™ Peptide Immobilization Kit via Maleimide | Peptide | -SH (Cys thiol) | Maleimide | Thiol ether | Stable |
SepSphere™ Peptide Immobilization Kit via Pyridyl Disulfide | Peptide | -SH (Cys thiol) | -S-S-Pyridine | Disulfide bond | Stable* |
SepSphere™ Peptide Immobilization Kit via Carboxylic Acid | Peptide | -NH2 | -COOH | Amide bond | Stable |
SepSphere™ Oligo Immobilization Kit via Carboxylic Acid | Oligo | -NH2 | -COOH | Amide bond | Stable |
SepSphere™ Oligo Immobilization Kit via Epoxide | Oligo | -NH2 | -CH(O)CH2 (epoxide) | β-amino alcohol | Stable |
SepSphere™ Oligo Immobilization Kit for Phosphate DNA | Oligo | -OPO3 (phosphate) | -COOH | Amide bond | Stable |
SepSphere™ Small Molecule Acid Immobilization Kit | Small molecule | -COOH | -NH2 | Amide bond | Stable |
SepSphere™ Small Molecule Amine Immobilization Kit via Glyoxyl | Small molecule | -NH2 | -C(=O)H (glyoxyl) | Secondary amine | Stable |
SepSphere™ Small Molecule Amine Immobilization Kit via Epoxide | Small molecule | -NH2 | -CH(O)CH2 (epoxide) | β-amino alcohol | Stable |
SepSphere™ Small Molecule Alcohol Immobilization Kit | Small molecule | -OH (aliphatic alcohol) | -NH2 | Thiol ether | Stable |
SepSphere™ Small Molecule Amine Immobilization Kit via Carboxylic Acid | Small molecule | -NH2 | -COOH | Amide bond | Stable |
SepSphere™ Small Molecule Ketone Immobilization Kit | Small molecule | -C=O- (Ketone or aldehyde) | -ONH2 (aminooxy) | Oxime bond | Stable* |
* Releasable under reducing conditions
The column in the SepSphere™ kit is the core component of molecular immobilization. The column consists of five parts.
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