Affinity Purification Mass Spectrometry (AP-MS)
Affinity purification mass spectrometry (AP-MS) is an effective method for the isolation and identification of binding partners to a target protein. The improvements in experimental workflows for antibody-based immunoaffinity purification (AP) of protein complexes as well as the technological progress in mass spectrometry (MS)-based proteomics, have greatly contributed to the unbiased characterization of virus-host protein interactions during the progression of a viral infection. Nowadays, the method has been successfully applied in multiple virology studies, including the isolation of virus-virus, virus-host multi-protein complexes, and the identification of direct and indirect protein interactions.
Affinity purification mass spectrometry (AP-MS)
AP-MS has emerged as an attractive high-throughput method for protein-protein interaction (PPI) study. During the process of AP-MS, a bait protein is selectively purified with specific antibodies and other affinity reagents that function as affinity capture probes for interacting from a cell or tissue lysate. Then the purified proteins are identified and quantified by MS. The experiments are then repeated with different baits to generate the combinations of bait-prey pairs, and protein networks are statistically analyzed on a computer. A major benefit of the AP-MS method is that it allows unbiased characterization of protein-protein interactions (PPIs) under physiological conditions. Moreover, the approach can provide quantitative information (q-AP-MS), which provides great convenience for identifying interaction partners and studying the influence of disturbance on PPI.
Service offering in Creative Proteomics
AP-MS has been employed to study virus-host protein interaction to provide temporal interaction events playing critical roles in different stages of a viral infection and to detect the overexpression of individual viral gene products to lead to an understanding of the functions of single proteins. We offer AP-MS techniques to help customers to achieve these goals. Since the composition of protein complexes of the protein is dynamic and highly affected by the time of day, tissue specificity, post-translational modification, and other conditions. In order to achieve successful AP-MS experiments, we focus on the specific project needs of our clients, experimental designs, and sample preparation.
Our services include the common workflow components, cell infection and disruption, immunoaffinity purification of protein complexes, digestion of proteins, and liquid chromatography coupled to mass spectrometry (LC-MS/MS), and bioinformatic analysis for protein identification. Among them, the preparation of samples is a crucial step. We offer cryogenic grinding using a ball mill, an effective and reproducible cell disruption method. The cryogenic cell lysis strategy before the immunoaffinity purification has been proved to help preserve protein complexes and weak protein interactions. And the cell disruption strategy is amenable to cell systems, tissues, and animal models for studying viral infections.
The AP-MS workflow.
Benefits of our services
- An optimization & scale-up platform
- Professional technical support
- A rapid and tailored solution for specific research applications
- Complete analytical study reports
To discover interaction events that are crucial for different stages of a viral infection, the characterization of virus-host protein interaction is essential. Creative Proteomics is a leading provider in the field of viral proteomics services. Based on the AP-MS method, we can help customers give insight into the progression and spread of a viral infection. For more information on how we can help you, please feel free to contact us. We look forward to providing services for your next project.
Reference
- Low, T. Y., et al. (2021). "Recent progress in mass spectrometry-based strategies for elucidating protein–protein interactions." Cellular and Molecular Life Sciences, 1-15.
* For research use only.
