In this application note, we use whey protein isolate and two casein macro peptides as an example to demonstrate the isolation and purification of bioactive peptide components with a system comprised of a puriFlash® 5.250 prepLC coupled online to an expression® CMS detector.
The new edition of our “Analytical Sciences, Synthesis & Purification” catalog is available now! Offering a full suite of innovative products and technologies from Advion Interchim Scientific, this catalog features over 20,000 products ranging from consumables and reagents to instrumentation.
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Q: WHAT IS THE FOCUS OF YOUR LAB’S RESEARCH?
A: The Rappsilber Laboratory is interested in studying the machinery of life, in particular protein structures and their interactions in living cells. Determining protein behaviour is crucial for developing new drugs or understanding terrible diseases such as cancer. We are doing this by developing state-of-the-art methods and new chemical tools for crosslinking mass spectrometry (crosslinking-MS), including new crosslinker reagents and crosslinking chemistry. Working on the chemistry-biology interface, our work encompasses chemical synthesis of peptides, fluorophore tagged sensors, peptide- and protein-reactive crosslinking reagents and bioorthogonal metal catalysts.
Q: WHAT WAS YOUR PREVIOUS WORKFLOW OR CHALLENGES?
A: This year (2020) we built a chemistry lab from scratch, here at the Institute of Biotechnology (TU Berlin). We had the opportunity to really think about our work flow requirements and create the most efficient chemical synthesis pipeline for a modern, 21st century, small chemistry laboratory. However, we are a satellite campus, located more than 10 km from the Institute of Chemistry, which meant that we were constrained by the lack of usual chemistry infrastructure. One of the major challenges that we faced here, is that without chemistry-specific analytical capability, we are working almost blind. Every step of chemical synthesis, including reaction monitoring, work-up, purification and final product characterisation required an exhausting and disruptive journey across Berlin!
Q: WHY DID YOU INCORPORATE THE EXPRESSION® CMS INTO YOUR LABORATORY?
A: We operate an advanced proteomics facility, with cutting edge, high-precision, high sensitivity mass spectrometers for analysing extremely complex peptide mixtures. But a chemist needs a workhorse, not a Formula 1 car. The Advion Interchim Scientific expression® CMS was a no-brainer. We needed a system that is versatile, compact, easy-to-use, easy-to-maintain (~10 minute calibration), robust and allows fast analysis (result in <30 seconds). This system is all of this, but furthermore it is a delight to use. The ASAP® Direct Analysis Probe works fantastically well for analysis of solids and liquids. Switching between APCI and ESI is so quick and easy. The Plate Express® system makes it a joy to identify reaction products directly from TLC plates, greatly simplifying subsequent purifications. The Plate Express™ also works really well for identifying synthesized peptides from TLC spots. The expression® CMS sits at the heart of our chemistry lab and workflow.
Q: WHO WOULD YOU RECOMMEND TO PURCHASE THE EXPRESSION® CMS?
A: We would recommend the Advion Interchim Scientific expression® CMS to literally any research laboratory, big or small. The versatility of the system is truly impressive.
The compound synthesis, purification and confirmation processes often present challenges. From ensuring successful synthesis of your product, setting up a flash purification method, and finally confirming ID of fractions, the lengthy work flow can be time consuming and repetitive.
This webinar will provide a foundation for flash purification processes, and will showcase a novel work flow concept that breaks down the process in to easy steps for optimal success. We will offer tips to optimize each essential segment of the workflow, and share seamless steps to quickly and easily drive the process from reaction monitoring to flash purification and fraction ID.
Presented by Dr. Daniel Eikel, Director of Customer Service and Product Applications, Advion.
This webinar was hosted by C&EN and recorded 8/13/20.
Mass spectrometry is an analytical technique used to determine the mass-to-charge ratio of ions in a sample and therefore the sample composition. This method is used across many industries including food and beverage, environmental monitoring, and clinical. There are many features of mass spectrometers that must be considered before purchasing such as the required mass analyzer technology, ionization source, and dissociation technique. Join Lab Manager and our panel of experts as we discuss how to decipher mass spectrometry technology offerings and find the right solution for your lab.
As an attendee, you will learn more about:
- Available technology for performing mass spectrometry
- Features to consider when purchasing mass spectrometry instruments
- How to find the right mass spectrometer for your application
Authors: Japan Bioindustry Association, Advanced Industrial Science and Technology, Japan
- Deacetylation of glycopeptide with an esterase prevents base-induced side reactions, b-elimination, and epimerization
- An esterase from B. subtilis was utilized for Deacetylation of sugar hydroxyl groups on glycopeptide synthesis
- Deacetylation with an esterase and glycosylations with glycosyltransferases were accomplished in one-pot
Glycopeptides are fragments of glycoproteins and are important in evaluating the biological roles of carbohydrates in glycoproteins. Fmoc solid-phase peptide synthesis using acetyl-protected glycosylated amino acids is a common strategy for the preparation of glycopeptides, but this approach normally requires chemical de-O-acetylation with a base that β-eliminates sugar residues and epimerizes the peptide backbone. Here we demonstrate a facile new chemoenzymatic synthetic strategy for glycopeptides, using an esterase for the de-O-acetylation of sugar residues and glycosyltransferases for successive sugar elongations at neutral pH.
ESI-MS spectra were collected using an Advion expression Compact Mass Spectrometer (CMS) in negative ion mode.
The Advion Interchim Scientific suite of systems and consumables allows users to harness the power of mass spectrometry, flash chromatography, prep LC and more. Simple and robust, download our brochure to learn more about:
- The Advion Interchim Scientific expression® compact mass spectrometer (CMS): A fast and easy analytical tool for the organic chemist. Ideal for fast reaction monitoring, the expression® CMS features a single quadrupole that can adapt to multiple ionization sources in seconds, including both ESI and APCI. The expression® CMS offers a variety of novel sampling techniques, including fast assay methods for liquids, solids, gases, and even air-sensitive compounds.
- Direct mass analysis of TLC plates in 30 seconds at the push of a button with Plate Express™
- One-touch analysis of solids and liquid samples with the ASAP® probe
- puriFlash® ultra performance flash purification: Ideal for method development and purification of rare and high added value compounds, the Interchim family of puriFlash® systems offer users a wide range of throughput options and the highest recovery rates at >95%.
- Mass-Guided Purification: the Interchim puriFlash® + Advion CMS offers the ideal solution for Flash-MS, and can provide fraction identification in <30 seconds.
Fill out the form to download the full Advion Interchim Scientific brochure now.
Advion Interchim Scientific’s Plate Express® is ideal for fast, easy TLC/MS analysis. Coupled with the expression® Compact Mass Spectrometer (CMS), the TLC plate reader allows prep-free sampling and data in less than 30 seconds.
Advion’s direct atmospheric solids analysis probe, otherwise known as ASAP®, permits chemists fast and reliable sample analysis of liquid and solid samples such as reaction mixtures, food samples, natural products and tablets.