Negative Control 2: Control VLP samples were stained with anti-Claudin18.2 antibody(Zolbetuximab biosililar IMAB362)at 1ug/mL, followed by Goat anti-human IgG Fc-PE secondary antibody. Negative Control 1: Claudin18.2 VLP samples were stained only with Goat anti-human IgG Fc-PE secondary antibody. From above data, the EC50 for IMAB362 binding with Claudin18.2 is 15.37ng/ml. Serial diluted Anti-Claudin18.2 monoclonal antibody (Zolbetuximab biosililar IMAB362) solutions were added, washed, and incubated with secondary antibody before ELISA reading. It can also be used as functional protein antigens to develop active antibodies with high drug potentials because the target protein on VLP exhibits a state like its native state on the cell surface.ĮLISA plates were pre-coated with 0.5ug/per well purified human Claudin 18.2 VLP. VLPs can be used for routine biochemical analysis, including ELISA, SPR affinity analysis, phage display screenings, protein labeling and cell binding experiments, Flow Virometry analysis, etc. The purified VLPs have the target MPs inserted in a complete bilayer phospholipid membrane structure, mimic the natural membrane-penetrating state of the protein (Figure 4). It is secreted from the surface of the cells that express target membrane proteins (MPs). Virus-like particles (VLPs) are self-assembling multi-protein nanoparticles with similar structural organization and conformation as viruses but without viral genome. Western blot of CCR8 membrane nanoparticles (MNPs) CCR8 full length membrane nanoparticles samples were stained with anti-CCR8 antibody (BME100063) at 2 μg/mL, followed by Goat anti-human IgG 488jsecondary antibody. Negative Control 3: CCR8 full length membrane nanoparticles samples were stained with anti-Claudin 18.2 antibody (an irrelevant antibody) at 2 μg/mL, followed by Goat anti-human IgG 488 secondary antibody.ĭ. Negative Control 2: Control membrane nanoparticles samples were stained with anti-CCR8 antibody (BME100063) at 2 μg/mL, followed by Goat anti-human IgG 488 secondary antibody.Ĭ. Negative Control 1: CCR8 full length membrane nanoparticles samples were stained only with Goat anti-human lgG 488 secondary antibody.ī. They can be used for the development of highly specific therapies targeting previously intractable targets such as ion channels and transporter.įigure 3:Membrane Nanoparticles (MNP) for the preparation of the full-length multipass transmembrane proteinsĪ. All platforms are based on the mammalian expression systems and prepare full-length membrane proteins in soluble protein products with native conformation and activity. To solve this issue, DIMA Biotech has developed several platforms for the full-length membrane proteins (MPs), including Membrane Nanoparticles (MNPs), exosome (EXO) and Virus-like particles (VLPs) et al (Figure 2). However, whole cell immunization often generate a lot of non-specific antibodies against other proteins, especially the abundant cytoplasmic proteins. It maintains the native conformation and modifications of target proteins, has high immunogenicity and doesn’t require complex purification steps. Whole cell immunization has been widely used for the mAb generation against transmembrane proteins. But they normally don’t have native conformation of target proteins, which can cause the loss of protein activities and some conformational epitopes. Extracellular domains (ECDs) are easy to be purified at high expression level. Each of them has shown its own benefits and issues (Table 1). To date, various antigens have been used for the preparation of mAbs against membrane protein (MP). Therefore, membrane proteins have been considered as “undruggable” targets in pharmaceutical companies. A good antigen is particularly critical for the development of monoclonal antibodies (mAbs) with therapeutic properties. Membrane proteins tend to lose the functions when removed from the membrane. Therefore, MPs account for more than 60% of all the current FDA-approved drug targets and 90% of the antibody drug targets.įigure 1: Membrane proteins play important roles in a variety of cellular functions.ĭespite the significance, drug discovery against membrane protein is still challenging, because it is difficult to find a good antigen for the generation of antibody against membrane proteins especially the multispan membrane proteins. Abnormal functions of MPs often lead to the occurrence of diseases. Membrane proteins (MPs) are the gatekeepers of cells and play an important role in a variety of cellular functions such as material transport, signal transduction, and cell-to-cell recognition (Figure 1). DiMPro TM Full Length Transmembrane Proteins
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