The Naveni® in situ proximity ligation technology makes it possible to visualize and quantify protein-protein interactions, post-translational modifications, and protein localization. The technology is based on dual antibody recognition and signal amplification to increase specificity and sensitivity.
Illuminating function in spatial proteomics
Understand biology – elucidate how proteins interact
Proteins perform a wide range of functions and are the closest way to understand biology. Proteins are also key targets in both health and disease. When proteins carry out their function, they interact with other proteins or molecules. To understand biological pathways and mode of action, we need to get a deeper insight into protein functions and how the protein interacts. Detecting the interactions has until now been either complicated, unspecific, or insensitive. The Naveni® Proximity Ligation Technology enables visualization of protein-protein interaction using one primary antibody for each target protein, without the need to disrupt the tissue microenvironment. Our newest technology design, Naveni® TriFlex concurrently visualizes two proteins in a free and complex state, enabling studies of protein interplay and functional state, not possible with any other technology.
Other common methods for studying protein-protein interactions, like BRET, cross-linking, pull-down assays, or co-immunoprecipitation (Co-IP), often require cell-modification or disruption of the tissue. This can lead to false-positive results in interaction studies since proteins that would not be in proximity in the intact tissue can be forced together in the lysate preparation or during cell processing.
The Naveni Proximity Ligation Technology enables new possibilities to:
- Understand biology and biological pathways
- Study protein functional states and interplay
- Identify new biomarkers at single-cell resolution
- Obtain deep phenotypic profiling in in vitro models, with early insight into the mode of action
- Demonstrate drug target engagement with local precision in tissues
- Distinguish biomarkers in tissue, with visualization of morphology and without disturbing the tissue microenvironment.
Read our application note on interacting glycoproteins in ovarian cancer (pdf)
Precise localization of your target with intact tissue morphology
Correct localization of proteins is critical for understanding and treating many diseases. The quality of in situ protein detection assays depends mainly on the specificity of the antibodies used against the target. Pathologists require intact cell and tissue morphology to confirm cancer cell type and origin of metastatic cancer; thus, enzyme-based chromogenic immunohistochemistry (IHC) of formalin-fixed paraffin-embedded tissue is considered to be the “gold standard” for clinical histopathology. But the quality of the IHC method relies largely on single antibodies. Many commercial antibodies have low specificity to the targets, generating unspecific background staining resulting in misleading results. The Naveni® Proximity Ligation Technology is based on dual antibody recognition, increasing the specificity leading to the correct local precision of the target.
Investigate PD1/PD-L1 interaction and activation in situ
PD1 PD-L1 signaling has with immune checkpoint inhibitors proven to be significant in cancer progression and treatment. Despite the recent success of immune checkpoint inhibitors, many patients do not benefit from these therapies, and predictive biomarkers improving patient stratification are needed. PD-L1 IHC is commonly used as a biomarker, but the correlation between PD-L1 expression levels and PD1/PD-L1 interaction is not always linear.
PD1 activation is an important step in the signaling pathway, yet phosphorylation of PD1 in situ is not well studied, mainly due to lack of specific methods.
Navinci has now developed two Proximity Ligation Assays for the specific detection of the PD1/PD-L1 interaction and phosphorylated PD1 in situ. The PD1/PD-L1 interaction has been verified in several tissues; Hodgkin’s lymphoma, Tonsil, Lung acinar adenocarcinoma, Lung squamous cell carcinoma, Malignant Melanoma, Colon adenocarcinoma and Pancreatic ductal carcinoma, see application examples here.
These products will give deep biological knowledge about the signaling pathways of PD1.
Precise localization of your target with intact tissue morphology
Correct localization of proteins is critical for understanding and treating many diseases. The quality of in situ protein detection assays depends mainly on the specificity of the antibodies used against the target. Pathologists require intact cell and tissue morphology to confirm cancer cell type and origin of metastatic cancer; thus, enzyme-based chromogenic immunohistochemistry (IHC) of formalin-fixed paraffin-embedded tissue is considered to be the “gold standard” for clinical histopathology. But the quality of the IHC method relies largely on single antibodies. Many commercial antibodies have low specificity to the targets, generating unspecific background staining resulting in misleading results. The Naveni™ Proximity Ligation Technology is based on dual antibody recognition, increasing the specificity leading to the correct local precision of the target.
Investigate PD1/PD-L1 interaction and activation in situ
PD1 PD-L1 signaling has with immune checkpoint inhibitors proven to be significant in cancer progression and treatment. Despite the recent success of immune checkpoint inhibitors, many patients do not benefit from these therapies, and predictive biomarkers improving patient stratification are needed. PD-L1 IHC is commonly used as a biomarker, but the correlation between PD-L1 expression levels and PD1/PD-L1 interaction is not always linear.
PD1 activation is an important step in the signaling pathway, yet phosphorylation of PD1 in situ is not well studied, mainly due to lack of specific methods.
Navinci has now developed two Proximity Ligation Assays for the specific detection of the PD1/PD-L1 interaction and phosphorylated PD1 in situ. The PD1/PD-L1 interaction has been verified in several tissues; Hodgkin’s lymphoma, Tonsil, Lung acinar adenocarcinoma, Lung squamous cell carcinoma, Malignant Melanoma, Colon adenocarcinoma and Pancreatic ductal carcinoma, see application examples here.
These products will give deep biological knowledge about the signaling pathways of PD1.
Unravel tumor progression - quantify phosphorylated receptor tyrosine kinases
Receptor tyrosine kinases (RTKs) are involved in cellular processes, and altered phosphorylation states may lead to disturbed signaling pathways, tumor progression, and cancer. To thoroughly investigate and elucidate the roots of tumor progression, it is crucial to detect and quantify phosphorylated RTKs. And specific tools for quantifying phosphorylation of RTKs in situ have been requested for a long time. Still, the lack of sensitive and specific commercially available assays has made quantification in situ problematic. The Naveni PTM pY product line makes it possible to quantify phosphorylated RTKs, HER2, EGFR, Met, and PDGFR-beta.
Our patented Naveni technology utilizes two Navenibodies, one with specific binding to the cytosolic C-terminal end of the RTK, and the other is pan-specific for phosphorylated tyrosine residues. Only when the two Navenibodies are in close proximity, do they produce an amplified signal leading to a quantifiable fluorescent dot with a high signal-to-noise ratio. By applying the Naveni® Proximity Ligation Technology, post-translational modifications (PTM) can be precisely detected to study the protein activation pathways.