Custom Membrane Protein Purification Service

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Overview Core Service Offerings Application Areas Project Workflow Case Studies Frequently Asked Questions (FAQs) Contact Us

Overview

Custom Membrane Protein Purification is a highly specialized service designed to provide customers with customized membrane protein purification solutions. Such services often involve the use of advanced technology platforms and multiple chromatography methods to achieve efficient expression and purification of target membrane proteins.

Specifically, Custom Membrane Protein Purification includes the following key steps:

Selection of cellular expression system: Select an appropriate cellular expression system based on the characteristics of the target membrane protein, such as mammalian cells, insect cells or bacterial systems.
Expression of recombinant protein: The gene of the target membrane protein is introduced into the host cell through genetic engineering to express it in the cell.
Separation and purification of membrane proteins: Different chromatography techniques (such as ion exchange chromatography, affinity chromatography, etc.) are used to separate and purify the expressed membrane proteins. These techniques can be used individually or in combination depending on the different characteristics of membrane proteins.
Purity and functional verification: Ensure that the purity and functionality of the final product meet customer needs through multiple analytical and characterization methods (such as mass spectrometry, dynamic light scattering, etc.).
Customized services: Provide personalized purification solutions based on customers 'specific needs to ensure that the final product can meet the requirements of specific applications.

This service is not only suitable for research, but is also widely used in drug development because membrane proteins are an important part of many drug targets. For example, G protein-coupled receptors (GPCRs) account for the majority of current drug targets.

Detergent-based phase separation can be used to purify or to concentrate membrane proteins(T Arnold, et al.,2007)

Core Service Offerings

As a CRO service outsourcing company, the core services provided by Custom Membrane Protein Purification include the following aspects:

Membrane protein expression: Expression of membrane proteins is a key step in the purification process of membrane proteins. Cell-based membrane protein expression methods are usually used, which is one of the most commonly used methods at present.
Membrane protein purification: Purification is an indispensable step in the preparation of membrane proteins. Because membrane proteins are different from water-soluble proteins, special care needs to be taken to avoid damaging their structure and function. Purification techniques for membrane proteins include the use of different detergents and buffers to remove impurities and further purify them through methods such as dialysis.
Membrane protein modifications: In some cases, membrane proteins may require specific chemical modifications to enhance their stability and biological activity. This includes but is not limited to modifications such as glycosylation, phosphorylation, etc.
Membrane protein characterization: In order to ensure the quality and function of membrane proteins, detailed characterization is required. This includes the use of various biophysical and biochemical techniques such as dynamic light scattering, mass spectrometry, X-ray crystallography, etc.
Process development and optimization: CRO companies usually provide optimization services for the entire production process, from genes to proteins, to ensure that every step achieves the best results.
Quality control: Including checkpoints such as protein purity, biological activity, batch-to-lot consistency, and low endotoxin levels.

Through these core services, CRO is able to provide customers with high-quality membrane protein purification solutions to meet their needs in drug research and development and biotechnology research.

Application Areas

Our company provides Custom Membrane Protein Purification service, which has extensive applications in the field of pharmaceutical research and development. The following are the application areas of the service and their detailed descriptions:

Drug discovery and development: Membrane proteins are more than 60% of current drug targets, so purification of membrane proteins is crucial to drug discovery and development. By optimizing the purification process of membrane proteins, the purity and stability of drug targets can be improved, thereby accelerating the development process of new drugs.
Structural biology research: Membrane proteins play an important role in cell membranes, and the study of their structure and function is very important for understanding biological processes and disease mechanisms. Through the purification of membrane proteins, high-quality samples can be provided for structural biology techniques such as freezing electron microscopy and X-ray crystallography, thereby revealing the three-dimensional structure of membrane proteins.
Biomedical research: Membrane proteins play a key role in various biological processes such as cell signaling, ion transport, and cell adhesion. Through the purification and functional study of membrane proteins, these processes can be deeply understood and new ideas for disease treatment can be provided.
In vitro diagnosis: Membrane proteins can be used as targets for antibody generation and diagnostic reagents. Through the purification of membrane proteins, high-quality antigens can be prepared, which can be used to develop diagnostic kits and improve the sensitivity and specificity of disease detection.
Vaccine development: Membrane proteins are expressed on the surface of certain viruses and bacteria and are important targets for vaccine development. Through the purification and functional study of membrane proteins, more effective vaccine candidates can be designed and the protective effectiveness of vaccines can be improved.
Gene therapy and cell therapy: Membrane proteins also have important applications in gene therapy and cell therapy. Through the purification of membrane proteins, high-quality therapeutic proteins or antibodies can be prepared for the design and implementation of gene therapy and cell therapy protocols.

Project Workflow

Our company's project workflow for providing Custom Membrane Protein Purification service is as follows

Membrane Protein Purification Project Workflow

Project launch: First, the customer submits membrane protein purification requirements, including sequence information and intended use of the target protein. The company formulates personalized project plans based on customer needs.
Cellular expression: Select appropriate host cells (such as Escherichia coli, insect cells, or mammalian cells) for expression of the target membrane protein. Use optimized expression systems to ensure high expression levels.
Membrane protein extraction: Use a specific membrane protein extraction kit (such as the Thermo Scientific Mem-PER Plus Kit) to separate membrane proteins from the cell membrane.
Dissolution and purification: Non-ionic detergents (such as DDM) are used to dissolve membrane proteins, and impurities are removed by methods such as ultracentular-centrifugation to maintain the natural conformation of the protein.
Optimization of purification process: Adjust detergent concentration and purification conditions based on the results of analytical ultracentrage (AUC) to obtain membrane proteins with high purity and low aggregation.
Purity testing and characterization: The purity and structure of purified membrane proteins were characterized using techniques such as high performance liquid chromatography (HPLC), mass spectrometry (MS) and electron microscopy (EM).
Functional analysis and application: Functional tests are carried out on the purified membrane proteins, such as enzyme activity determination, drug screening, etc., to verify their biological functions.
Customer delivery: Deliver purified and strictly quality controlled membrane proteins to customers, and provide detailed experimental reports and data support.

Through the above processes, we are committed to providing customers with efficient and reliable membrane protein purification services to help customers achieve success in biomedical research and drug development.

Case Studies

Background: TmrA is an important transmembrane protein widely used in the study of drug efflux and intracellular transport. However, its purification process has always faced challenges, especially in maintaining protein homogeneity and reducing aggregation.

Solution: We used analytical ultracentultracentular-centrifugation (AUC) technology to optimize the purification process of TmrA. By comparing the effects of different detergents (such as OG, Triton X-100, and DDM) and their concentrations on the behavior of TmrA, we found that DDM was more effective in purifying TmrA at high concentrations, and supplementing DDM at low concentrations maintained protein homogeneity and aggregation.

Result: Through the optimized purification process, we successfully obtained high-purity, uniform and non-aggregation TmrA protein, providing high-quality samples for subsequent structural biology and biomedical research.

Frequently Asked Questions (FAQs)

In order to help customers better understand the Custom Membrane Protein Purification service provided by our company, the following are common questions and answers:

1. What is membrane protein purification?

Answer: Membrane protein purification refers to the process of extracting and separating membrane proteins from cell membranes. Because membrane proteins are hydrophobic, detergents are needed to stabilize these proteins during the purification process.

2. What is the importance of membrane protein purification?

Answer: Membrane proteins play an important role in cellular functions, such as regulating ion transport across the membrane, transferring chemical or electrical signals, etc. Since membrane proteins are the main body of many drug targets, their purification is crucial for biomedical research.

3. What detergents are commonly used when purifying membrane proteins?

Answer: Commonly used detergents include but are not limited to alkyl maltosides (such as β-DDM) and glucosides (such as octyl β-D-glucopyranoside). Studies have shown that DDM is better at purifying certain membrane proteins such as TmrA.

4. How to overcome the hydrophobicity of membrane proteins?

Answer: A common method is to use molecular biology technology to build a recombinant membrane protein containing fusion affinity tags, so that the target protein can be more easily modified and manipulated.

5. How to remove detergents during the purification process?

Answer: Removal of detergents is usually carried out through methods such as dialysis or ultrafiltration. Dialysis is a commonly used method to remove excess detergent by exchanging a sample with a buffer with a lower detergent concentration.

6. How can purified membrane proteins be used for structural studies?

Answer: The purified membrane protein can be structurally studied by cryogenic electron microscopy. First, it is necessary to obtain a purified membrane protein sample wrapped in a detergent, then dissolve the appropriate phospholipid molecule in the corresponding detergent, and then perform subsequent structural analysis.

Contact Us

Our membrane protein purification process uses a variety of advanced technologies, including but not limited to liposomes, nanodisks, and virus-like particles. We use optimized detergent conditions and ultracentular-centrifugation technology to ensure the stability and functionality of membrane proteins during the purification process.

If you need to learn more about our services or have any questions, please feel free to contact us. We look forward to working with you to jointly promote the progress of scientific research.

Please note that all services are for research use only. Not intended for any clinical use.

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