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HighlightsWith over a decade of experience in phage display technology, Creative Biolabs can provide a series of antibody or peptide libraries that are available for licensing or direct screening. These ready-to-use libraries are invaluable resources for isolating target-specific binders for various research, diagnostic or therapeutic applications.
HighlightsCreative Biolabs has established a broad range of platforms for developing novel antibodies or equivalents. These cutting-edge technologies enable our scientists to meet your demands from different aspects and tailor the most appropriate solution that contributes to the success of your projects.
HighlightsWith deep understanding in antibody-related realms and extensive project experience, Creative Biolabs offers a variety of references to help you learn more about our capacities and achievements, including infographic, flyer, case study, peer-reviewed publications, and all kinds of knowledge that can assist your projects. You are also welcome to contact us directly for more specific solutions.
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HighlightsProkaryotic ubiquitin-like protein (Pup) is a functional analogue of ubiquitin found in prokaryotic Mycobacterium tuberculosis and is the only known post translational modification protein modifier in prokaryotes to date. Pupylation is a form of protein post translational modification (PTM) evolved by Mycobacterium and other actinomycetes. The study of pupylation has important implications for the treatment of Mycobacterium tuberculosis infections, in addition to providing insight into the mechanism of action of pupylation. Based on the phage display platform and hybridoma platform, Creative Biolabs provides a comprehensive pupylation-specific antibody development service to facilitate the progress of pupylation research.
Unlike eukaryotes, prokaryotes lack well-defined subcellular compartments and therefore have additional requirements for specificity and regulation of protein hydrolysis. Pupylation is a post-translational modification of the lysine side chain with a Pup that targets proteins for proteasomal degradation in Mycobacterium and other members of the actinomycetes. After Dop deamidation, Pup is ligated to the substrate Lys via an isopeptide attached to the C-terminal carboxylate of Glu. For simplicity, the ligation is to the γ-carboxylate. Pup interacts with the AAA-ATPase Mpa, which may unfold the substrate and feed it into the catalytic chamber of the proteasomal core for degradation. It is not known whether additional factors are required for optimal binding and delivery to the proteasome, whether PafA and Dop interact with the proteasome, or whether Pup is recycled or degraded. During pupylation, Pup is covalently linked to the target protein by the formation of an isopeptide bond between the γ-carboxylate of its C-terminal glutamate and the lysine side chain in the target protein. Pup is the only known prokaryotic protein that is functionally similar to ubiquitin. Although functionally homologous, Pup differs from ubiquitin in many other ways. With little sequence and no structural homology to ubiquitin, Pup is first deamidated by Dop and subsequently coupled to a variety of substrates via PafA. The two proteins are highly similar to each other but have no similarity to eukaryotic ubiquitin/proteasome-related proteins. Pupylation is a reversible process. Following the Dop-catalyzed deamidation reaction, Pup binds to the substrate (e.g. FabD) via an isopeptide linkage between one of the C-terminal side chain carboxylates of the substrate, Lys, and Glu, or a C-terminal backbone carboxylate. The ligation reaction is catalyzed by PafA and is ATP-dependent. For simplicity, ligation is performed via the γ-carboxylate of Glu. And depupylation can be performed by hydrolysis, transamidation or other means of depeptidase reaction.
The development of pupylation-specific antibodies is not an easy task. Different antigens with varying modification sites, diverse immunized animals, and various immunization procedures can all impact the obtained antibodies and, consequently, the final experimental results. Therefore, in general, selecting regions with highly conserved and unique properties as antigens can enhance the specificity and affinity of antibodies. In addition, considering that Pup proteins may undergo conformational changes during the modification process, it is important to select antigens that reflect their natural state. When selecting immune animals, the immune response to Pup proteins or peptides may vary among different animals, making the selection of suitable immune animals a factor that affects the quality and yield of antibodies. After obtaining pupylation-specific antibodies, it is necessary to test the specificity and cross-reactivity of the antibodies to determine their potency.
Creative Biolabs has a wealth of knowledge and experience in PTM specific antibody discovery. We would be happy to discuss our knowledge and experience in pupylation-specific antibody development with you.
All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.
