East Port Seminar: Efficient Tools for MS Proteomics (Promega, JPT Technologies)

We would like to invite you to a seminar that we organize in cooperation with the German company JPT Technologies. This seminar will focus on MS proteomics and it will introduce tools for efficient sample preparation and quantification provided by Promega and JPT Technologies. The seminar will be again split into two parts.

Part 1 – Sample preparation tools for robust, reliable, and accurate mass spec protein analysis

Protein sample preparation for mass spectrometry analysis is a complex process with many pitfalls. This part of the seminar will cover solutions for common shortcomings of protein mass spec sample preparation and discuss the advantages of Promega tools and reagents that can help achieve robust, reliable, and accurate protein and proteome characterization with mass spectrometry. We will review the application of our tools and reagents to improve protein digestion and biotherapeutic protein characterization with peptide mapping.

In this part you will learn the following:

• How to achieve efficient protein extraction, solubilization, and digestion
• How to avoid unwanted side effects such as non-specific cleavages, non-enzymatic protein modification, and extensive trypsin autoproteolysis
• How to complete the entire protein sample preparation in-gel and in-solution within 1h
• How to increase protein sequence coverage using novel proteases

Speaker:

Vojtěch Ledvina, Ph.D. – application specialist, East Port Praha

Part 2 –  Simple robust protein quantification using the SpikeTides peptides (JPT Technologies)

Proteomics is the study of a cell's protein inventory at different times by protein identification and quantification. The application of LC-MS/MS has tremendously facilitated this process. Generally, samples containing relevant proteins are digested into peptides and analyzed by LC-MS/MS as a surrogate measurement of protein levels. To increase the accuracy of detection and to enable protein quantitation, chemically synthesized stable isotope-labeled (SIL) peptides can be added to the sample. JPT's peptide synthesis technology enables the ultra-fast and inexpensive provision of light or heavy isotope stably labeled reference peptides and peptide pools for proteome-wide profiling by mass spectrometry. SpikeTides™ reference peptides and SpikeMix™ peptide pools enable robust, multiplexed identification and relative quantification of protein expression levels.

Whilst SpikeTides TQL with the proprietary QTag, a small chemical tag attached to each peptide, allowing robust and reproducible quantitation via HPLC-UV or HPLC-MS and UV without the need to go via traditional cumbersome and costly quantification methods such as AAA. The QTag is readily released by digestion with trypsin and does not interfere with subsequent measurements. The method is very accurate and has been validated in numerous peer-reviewed papers.

During this talk you will learn about the following:

• Peptide synthesis and quality levels
• Human proteome atlas
• Different peptide formats for targeted and functional proteomics
• Methods for quantification and Qtag peptides as alternatives to absolutely quantified peptides
• Different Kits for standardization
• References for PTM & Retention time standardization
• Application examples such as immunopeptidomics

Speaker:

Michael Drosch, Ph.D. – JPT Technologies

Harmonogram:

9:30-10:00    Registration
10:00-11:00   Sample preparation tools for robust, reliable, and accurate mass spec protein analysis (Promega)
11:00-12:00   Simple robust protein quantification using the SpikeTides peptides (JPT Technologies)

For whom is the webinar intended?

The seminar is open to all interested parties regardless of their field of study and age and is also suitable for university students. Registration is required for participation. The seminar will be conducted in English.