Antibody and Vector Core

University Of Maryland Baltimore, Baltimore MD

Investigators

Linked publications & trials

Abstract

Antibody and Vector Core: Project Summary Over the past 15 years, the Maryland PKD Research Core Center (MPKD-RCC) Antibody and Vector Core has served as the nation’s primary resource for basic research tools, including antibodies and vectors for PKD proteins (Polycystin-1, Polycystin-2, and Fibrocystin), to advance PKD research. The overarching goal of the Core is to develop and distribute innovative, validated PKD antibodies and vectors, along with technical support, to foster the next generation of discoveries in the field. The field currently faces a critical shortage of these tools, particularly high-quality antibodies for in vivo studies, which poses a significant barrier to progress. Studying PC1 is especially challenging due to its low abundance, large size, multi-transmembrane structure, and the biochemical complexity introduced by GPS cleavage, resulting in the N-terminal fragment (NTF) and the 11-transmembrane C-terminal fragment (CTF). Additionally, the large size of these proteins complicates the manipulation of PKD vectors. This application leverages the Core Director’s decades of expertise in polycystins and fibrocystin and his steadfast commitment to providing technical support to the PKD research community. In this funding period, we propose the following specific aims: Aim 1. Develop novel rabbit monoclonal antibodies against human PC1-CTF using RNA immunization. Aim 2. Develop novel rabbit monoclonal antibodies against mouse PC1 NTF using mammalian-expressed mouse NTF immunogen. Aim 3. Develop novel rabbit monoclonal antibodies against human PC2 using PC2 immunogen produced in mammalian cells. Aim 4. Maintain and provide the current PKD molecular resources. Aim 5: Provide technical consultation and assistance with experimental design, troubleshooting, and data interpretation. This project anticipates generating transformative rabbit monoclonal antibodies for comprehensive in vivo analyses of polycystins across a wide range of applications, filling critical gaps in the availability of high-quality antibodies in the field. Notably, these new immunoreagents will enable super-resolution microscopy to precisely localize polycystins and analyze the spatial relationship between NTF and CTF, as well as between PC1 and PC2. Additionally, testing these antibodies for modulating PC2 and PC1/PC2 channel functions represents a cutting-edge functional application, advancing the frontiers of PKD research. In summary, the Antibody and Vector Core will generate high-quality rabbit monoclonal antibodies against human PC1-CTF, mouse PC1-NTF, and human PC2, expanding the available toolkit for a variety of demanding applications. We will also maintain access to existing PKD molecular resources and provide expert guidance to ensure effective use of both current and new tools, facilitating and accelerating discoveries in PKD research.