Fully Automated in situ Proximity Ligation Assays from Navinci Diagnostics

For the first time, a proximity ligation assay with a fully automated workflow is commercially available. With the NaveniBright™ and the Naveni® PD1/PD-L1 BOND RX HRP, our partner Navinci Diagnostics has introduced two new groundbreaking products that enable seamless integration of chromogenic readout automation on Leica Biosystems' fully automated research stainer BOND RX/RX^m. The kits have been carefully developed based on the innovative Naveni® in situ proximity ligation technology. With this portfolio extension, automated detection of protein-protein interactions and post-translational modifications in situ is possible for the first time. 

These topics await you:

1) Automated Detection of Protein-Protein Interactions and Post-translational Modifications in situ

2) Naveni® Kits for BOND RX/RX^m

3) Benefits and Validation of Naveni® Kits for BOND RX/RX^m

4) What is needed to run NaveniBright™ BOND RX HRP and/or PD1/PD-L1?

5) Investigating Protein-Protein Interactions for the Understanding of Protein Function and Signaling Pathways

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Automated Detection of Protein-Protein Interactions and Post-translational Modifications in situ

With the new kits from Navinci Diagnostics for the Leica BOND RX/RX^m, you can efficiently detect protein-protein interactions and post-translational modifications with high precision and accuracy. Visualize specific protein targets such as PD1/PD-L1 while maintaining intact cell and tissue morphology for a holistic understanding of spatial protein relationships. A seamlessly integrated, automated workflow on BOND RX/RX^m also improves your efficiency and reproducibility, optimizes the examination process and achieves high throughput. Save valuable time with the automated workflow (see below), reduce time spent and maximize overall research productivity. The kits are optimized for FFPE tissue and the reagents included are sufficient for 30 FFPE tissue sections.

Naveni® Kits for BOND RX/RX^m

NaveniBright™ BOND RX HRP

NaveniBright™ BOND RX HRP offers flexibility tailored to your unique primary antibodies and targets. Use your own primary antibodies validated for IHC on BOND RX/RX^m, add NaveniBright™ BOND RX HRP for proximity ligation assays and illuminate the function of your protein targets.

View NaveniBright™ BOND RX HRP

Naveni® PD1/PD-L1 BOND RX HRP

Naveni® PD1/PD-L1 BOND RX HRP precisely visualizes the interaction of PD1/PD-L1, providing a comprehensive solution for studying these interactions in the tissue microenvironment (Fig. 1). The kit provides researchers with detailed spatial insights into the dynamics of immune checkpoint signaling pathways and can facilitate the development of targeted immunotherapies for cancer treatment. The primary antibodies are optimized and already included in the kit.

View Naveni® PD1/PD-L1 BOND RX HRP

Figure 1: Naveni® PD1/PD-L1 BOND RX HRP was used to detect PD1/PD-L1 interaction in head and neck cancer (left, center), the adrenal gland served as a negative biological control (right).

Benefits and Validation of Naveni® Kits for BOND RX/RX^m

If you use the new Naveni® kits for BOND RX/RX^m, you will benefit from numerous advantages:

• Enhanced Research Insights: Clear visualization of protein interactions and post-translational modifications in tissue microenvironments
• Improved Efficiency and Reproducibility: Automated assay workflows reduce hands-on time and minimize variability
• Superior Productivity: Time-saving automation and streamlined workflows maximize research productivity enabling high throughput studies using proximity ligation assay.
• FFPE optimized for 30 tissue slides
• Validated on FFPE tissue and cell samples
• Standard Template in Leica BOND software

In addition, the kits have undergone thorough validation. NaveniBright™ BOND RX HRP has been validated on a variety of FFPE tissues and in five different interaction assays (PD1/PD-L1, CD8/MHC-I, mesothelin/mucin, E-cadherin/beta-catenin and Zap70/Lat) as well as a dual recognition assay (HER2/HER2). Naveni® PD1/PD-L1 BOND RX HRP has also been validated in various FFPE tissues, lung cancer, ovarian cancer, squamous cell carcinoma of the head and neck, lymph nodes and tonsils. Adrenal glands or PD-L1-positive cell lines were used as biological negative controls.

The validation protocol included a thorough, comparative analysis between the automated kits and the NaveniBright™ HRP and Naveni® PD1/PD-L1 HRP using the manual methodology (Fig. 2). Of note, the kits were implemented at three different research sites, each utilizing their own BOND RX/RXm systems to ensure the establishment of robust performance characteristics in different experimental environments.

Figure 2: The automated method leads to the same results as the manual method; further comparisons can be found in the application note "High-throughput fully automated in situ proximity ligation assays for spatial interactomics".

What is needed to run NaveniBright™ BOND RX HRP and/or PD1/PD-L1?

In addition to the respective kits from Navinci Diagnostics, you will also need other reagents and tools, primarily from Leica, as well as the corresponding software. The protocol for the automated Navinci kits is called "NaveniBright". Leica BOND RX/RX^m already in operation will be automatically updated with the new software, while newly purchased Leica BOND instruments will be supplied with the latest software. The required reagents are listed below:  

Leica BOND Detection Systems and Containers:

• BOND Titration Kit (OPT9049)
• BOND Titration Inserts (OPT9719)
• BOND Intense R Detection (DS9263) oder BOND Polymer Refine Detection (DS9800)

Leica Bulk Reagents:

• BOND Wash 10x Concentrate (AR9590)
• BOND Dewax (AR9222)
• BOND Epitope Retrieval Solution 1 (AR9961)
• BOND Epitope Retrieval Solution 2 (AR9640)
• Reagent Grade Alcohol
• De-ionized water
• BOND Universal Covertiles (S21.4611)

Please note that you still have to purchase these reagents from Leica Biosystems, Navinci does not offer these products. For post-processing and mounting, you will also need water, isopropanol, mounting medium and coverslips.

Investigating Protein-Protein Interactions for the Understanding of Protein Function and Signaling Pathways

The Naveni® kits for the Leica BOND RX/RX^m fully automated research stainer enable researchers to study complex protein-protein interactions with high precision and accuracy using their own primary antibodies. The kits allow researchers to study protein functions and signaling events in the tissue microenvironment. By visualizing protein-protein interactions in the spatial context of cells and tissues, intricate signaling networks can be deciphered and new therapeutic targets for various diseases can be uncovered.

One example is the interaction between CD8 and MHC-I (Fig. 3). Understanding these interactions is crucial to clarify the role of cytotoxic T cells in immune surveillance and their involvement in diseases such as cancer and autoimmune diseases. Another example is the dynamic interaction between E-cadherin and beta-catenin. It provides insights into cell adhesion and signaling pathways that are critical for understanding tissue organization and progression of various diseases, including carcinoid tumors.

Figure 3: Staining of CD8/MHC-I interaction in breast cancer with NaveniBright™ BOND RX HRP (A, B: positive signals, C: negative control).

Through the collaboration between Leica Biosystems and our partner Navinci Diagnostics, you will benefit from Navinci's expertise in the accurate study of protein interactions in the tissue microenvironment and Leica's renowned automation capabilities. The two fully automated Naveni® kits for BOND RX/RX^m will standardize and simplify the assay workflow for all laboratories and researchers. These new assay solutions will provide the scientific community with groundbreaking tools to advance research and significantly improve patient care.

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