Membrane Protein Preparation as Antigen for Antibody Discovery
Empowered by plenty of cutting-edge platforms and technologies, Creative Biolabs has long been committed to high-quality membrane protein antibody discovery services. With more than 10 years’ experience, we are dedicated to helping our clients prepare membrane protein as antigens in various formats for antibody generation & selection.
Since their vital physiological functions, membrane proteins are popular drug targets for therapeutic and diagnostic applications. But the intricate structures and low natural abundance make most membrane proteins very difficult to express and isolate. How to obtain membrane proteins in their stable, native and functional format poses a major challenge for most relevant research programs. In particular, discovery of anti-membrane antibodies is often lagged due to the difficult antigen preparation process. In aim of overcoming this issue, Creative Biolabs has established a versatile membrane protein technique platform providing multiple strategies, from which you can always find a best fit for your specific target:
Anti-Membrane Antibody Production by DNA Immunization: this is one of the most effective methods to circumvent tedious membrane protein preparation. With the help of DNA immunization, the DNA of target protein can be directly introduced to host cells, resulting in spontaneous protein expression in native structures and native membrane environments. Meanwhile, it has minimal cytotoxicity to a wide range of hosts. It’s also very simple-to-operate, so different immunogen designs can be easily tested.
Anti-Membrane Antibody Production Using Recombinant Proteins: this is the most widely used approach to prepare membrane proteins antigens. Based on our world-leading cell-based expression and cell-free expression systems (prokaryotic/eukaryotic), Creative Biolabs offers professional membrane protein expression and purification services from small to large scales upon request. Aided by our detergent screening platform & lipid screening platform, highly purified membrane proteins can be reconstituted into multiple formats, such as detergent micelles, liposomes, nanodisc and polymers. Besides, lipoparticles, a special type of purified virus-like particles (VLPs), are another format to present integral membrane protein as immunogens. Well-designed peptides can also be utilized as surrogate of full-length membrane proteins with similar immunogenicity and characteristics but with higher stability and solubility.
Anti-Membrane Antibody Production by Whole Cell Antigen Presentation: both transient transfection and stable transfection can be used as antigens overexpressing membrane proteins presented on native cell membrane with native conformations and modifications. Creative Biolabs is proficient in constructing stable cell lines with membrane protein overexpressing from CHO, HEK, dendritic cells, etc. With years of efforts, we have tailored over 200 GPCR/ion channel stable cell lines with high expression levels. They can be employed directly for membrane protein isolation or host immunization. Plus, antigens presented on dendritic cells show higher immunogenic effect.
Anti-Membrane Antibody Production by Exosomes: as secreted membrane nanovesicles, exosomes have small size with the diameter range from 50-100 nm. The orientation of membrane proteins on exosomes is close to the orientation of the whole cell. Besides, exosomes can be acquired from numerous cell types and have the similar ability with their source cells. They have stable storage property and can cause strong immune response without the additional adjuvant.
With the help of this cutting-edge platorm, Creative Biolabs has successfully obtained many functional GPCR immunogens, ion channel immunogens and transporter immunogens using multiple approaches. They can serve as excellent antigens for raising high specific, high affinity antibodies by means of phage display, hybridomas, B cell sorting technology, etc. We are more than happy to share our experience and help our customers to solve all kinds of challenges in this regard. Please feel free to contact us for more details.