Affinity Purification Services

Protein purification is the basic technology of biological research and industrial production, and the purification method with high efficiency, convenience, economy and environmental protection has always been pursued by people. Currently, affinity purification is the most selective and versatile form of liquid chromatography, relying on reversible and specific binding between proteins and their associated ligands. For example: the binding of hormones to their receptors, the interaction of enzymes with their receptors, and the binding of antibodies to their target antigens. The specificity of this interaction is achieved by immobilizing an interacting agent (affinity ligand) on the column as a stationary phase support, and the target protein is eluted from the complex mixture through selective absorption of the affinity ligand. A complex mixture containing the target protein is applied to a mobile phase of suitable composition and pH to allow the target protein to bind to the affinity ligand while other components pass through, and the retained target is then eluted from the column.

 

The key to affinity purification services is to select appropriate affinity ligands. Affinity ligands can include antibodies, small molecules, nucleic acids, or proteins and peptides that specifically bind to the target protein. Affinity protein purification is a crucial step in protein research, leveraging the affinity between proteins and specific molecules (e.g., antibodies, ligands, or metal ions) to achieve selective isolation and purification of target proteins. Affinity purification services provide an effective means to purify and identify specific proteins from complex biological samples, as well as to analyze their physicochemical properties and biological functions. This enables researchers to better understand fundamental mechanisms in life processes and the onset and progression of diseases.

 

Affinity chromatography utilizes various types of binding agents (ligands) and stationary supports (matrices). By optimizing the properties of these components, such as specificity, selectivity, reproducibility, chemical reactivity, and cost-effectiveness, this method can be applied to large-scale protein purification to achieve the desired yield and purity. Over the years, the scientific community has made significant efforts to find or manufacture high-quality affinity ligands and matrices to aid in the development of efficient affinity chromatography methods aimed at increasing the purity and yield of desired proteins in a cost-effective manner.

 



Xue, A. et al. Biomolecules. 2022.

Figure 1. Chromatography Matrix Used for Antibody Affinity Purification.

 

Services at MtoZ Biolabs

Based on advanced technical analysis platforms, the affinity purification services provided by MtoZ Biolabs can obtain purified proteins efficiently, simply and quickly.

 

Service Advantages

1. Advanced Analytical Platforms

MtoZ Biolabs has established advanced affinity purification services platform, ensuring reliable, fast, and precise purification processes.

 

2. Transparent Pricing

Our pricing is transparent, with no hidden fees or additional fees.

 

3. Custom Research Solutions

MtoZ Biolabs offers tailored services to address your unique research challenges and experimental requirements.

 

Applications

1. Ni Column Purification

Nickel ions in Ni columns bind to alkaline proteins tagged with six histidine residues.

 

2. GST Agarose Purification

Purification of glutathione S-transferase (GST) fusion protein through protein affinity purification services can obtain high-purity GST fusion target protein in one step, with mild purification conditions to ensure protein activity.

 

3. Protein A

Protein A is commonly used for the purification and separation of IgG. It is widely applied to isolate and purify various types of IgG or IgG fragments from biological fluids or cell culture supernatants.

 

4. Strep-tag II Purification

Affinity purification techniques are employed for protein expression and purification in systems such as prokaryotic and mammalian cell expression systems.

 

Case Study

1. Cationic Affinity Purification of Histidine-tagged Proteins

Various non-covalent interactions exist within and between proteins, including hydrogen bonding, electrostatic interactions, hydrophobic interactions, and coordination bonds. These interactions form the foundation of most protein purification methods, some of which involve salts. Alkaline earth cations (e.g., Mg²⁺, Ca²⁺), alkali metal cations (e.g., Li⁺, Na⁺, K⁺), and even non-metal cations (e.g., NH₄⁺) can precipitate histidine-tagged proteins at low salt concentrations. These precipitated proteins can dissolve in solutions with appropriate ion concentrations. Based on this principle, a novel cationic affinity purification method was developed, achieving highly purified proteins after only three centrifugation steps. The purification yield is comparable to that of immobilized metal affinity chromatography. The interaction between histidine-tagged proteins and cations holds significant potential for broader applications.

 



Hong, X S. et al. Applied Microbiology and Biotechnology, 2023.

Figure 2. Schematic Representation of Cation-Induced Precipitation and Dissolution of Histidine-Tagged Proteins (with Mg²⁺-Containing ND).

 

Deliverables

1. Protein Purification Results

 

2. Experimental Records

 

3. Raw Data Files

 

4. Experimental Report

MtoZ Biolabs, an integrated Chromatography and Mass Spectrometry (MS) Services Provider, provides advanced proteomics, metabolomics, and biopharmaceutical analysis services to researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our ultimate aim is to provide more rapid, high-throughput, and cost-effective analysis, with exceptional data quality and minimal sample consumption. Free project evaluation, welcome to learn more details!