• Molecular Diagnostics
• RNAi
• Screening

The event was hosted by the Genome Institute of Singapore located in the rather lovely environment of the Biopolis site at the One-North site in Singapore. One-North is a research district set up by the Singapore government and this site is dedicated to biomedical research. It consists of several dedicated buildings all linked by sky bridges to symbolise the networking between research areas. Institutes based here are overseen by The Agency for Science, Technology and Research (A*Star) and pharma companies such as the  Novartis Institute of Tropical Disease is also located on the site. The site is internationally renowned and has scientists from all over the world (over 2500 scientists at present). Whilst here we had the chance to meet up with an old friend at the Experimental Therapeutics Centre. The ETC was set up in 2007 to advance and develop drug discovery in Singapore specifically to take forward promising research from academia into proof of concept and first in human trials. It has an interesting remit and looks at peptidomimetics and new antibody technologies so is at the forefront of our field.

The meeting itself was disappointingly quiet despite the great location; considerably quieter than the previous SelectBioscience molecular diagnostics meeting in San Francisco. The focus was very much on RNAi and diagnostics did not feature heavily in the content; there were only 3 diagnostics posters presented at the conference! As a result our custom peptide synthesis and custom antibody services were not in great demand. More interest was expressed in Peptide Nucleic Acids (PNA) which we distribute for Panagene, but unfortunately not in this area of the world.

This meeting did present a good opportunity to finally visit Singapore and continue CRB’s expansion into Asia but we would be unlikely to attend this one again. The research community in Singapore seems to be vibrant despite this and we hope to be working with more of you soon. Having supplied customers in the area for many years we know it is very easy to ship our custom materials here and as Singapore itself speaks English this plays to our strengths in custom reagent generation. The key to great reagents is in understanding the customer end requirements and this is something we strive to achieve in every case; a common language certainly helps with this! As the UK and Singapore have such a shared history we would like to build upon this by collaborating with researchers here. If you are a Researcher in Singapore please do get in touch to see how we can help.

The location of the meeting was the South San Francisco Conference Centre and the venue provided to be very good with excellent coffee and meals provided during those vital breaks where all the real thinking and science gets done. The layout was very good and the conference organisers had done a good job of integrating exhibitors with delegates whilst managing to accomodate a large number of poster boards presenting some interesting science.

We met a variety of people from all scientific fields as the attendees from all the threads met and mingled in the exhibition hall. Scientists had come from all over the world for this meeting and we had the opportunity to discuss the Rugby World Cup (and a bit of peptide chemistry!) with scientists from Australia and New Zealand. Scientists were present from all areas of the scientific community; academia, traditional big pharma, biotech and diagnostics companies were all represented and this mix made for some varied and interesting presentations and discussions.

The big area of discussion for us seemed to be the use of polyclonal or monoclonal antibodies within flow diagnostics and lab-on-a-chip applications i.e. capture antibodies within chip based screening or cell sorting type devices. This is an interesting and constantly developing area and it seems that as with all areas in science the interface between Physics and Biology is producing some good collaborations, research and products. This is of course,  very tied into the field of personalised medicine, which is a current buzzword and area of great interest for the healthcare community.

From our perspective this was a very valuable meeting and we had interest in custom peptides, custom antibodies, fluorescent labelling and PNA (Peptide Nucleic Acids) so we were kept busy answering questions; some easy and some a little more challenging which is appreciated as for those of us who left the lab some time ago it is nice to get into some real science.

It would be nice to attend this meeting next year, we will certainly be considering it and we look forward to meeting many more of you at future meetings.

What we liked:

• The friendly atmosphere: the peptide world is a small world and the APS meeting is a fantastic opportunity to network and catch up with friends
• The quality of the speakers (some very distinguished)
•  Jennifer Ottesen’s² synthesis of acetylated Histone H3 using a Dawson Dbz resin with alloc protecting groups (to avoid the nasty peptide branching)
•  A lot of Click chemistry
•  Jean Chmielewski’s³ functionalised collagen to produce spheres, discs or fibers
•  Christian Becker’s⁴ optimised synthesis of a protein with 3 transmembrane domains.
•  San Diego

What we didn’t like:

•  The packed lunches! Nothing memorable…
•   The tent, so far from the lecture hall.

Useful links:

¹ Ashaf Brik’s lab at Ben Gurion University
² Jennifer Ottesen’s lab at Ohio State University
³ Jean Chmielewski’s lab at Purdue University
⁴ Christian Becker – Center for Integrated Protein Science Munich 

This meeting brought together over 17,500 people from all walks of life with an interest in Diabetes from research through diagnostics and medical to patients so there was an interesting mix of people browsing the exhibition and some interesting sights to see. All of us at CRB joined in the fray and took advantage of the free haemoglobin A1C tests being provided by suppliers such as Biorad and Novo Nordisk. A1C is a biomarker of interest in determining if a person is pre-diabetic or diabetic. I am pleased to say that none of us were surprised with a positive for diabetes result but it did illustrate the growing importance of biomarkers and personalised medicine. Increasingly we find that our custom antibody customers are working within the field of  biomarker discovery and within the diagnostics field so it was a good venue for us to see what this all leads to in the “real” world. Traditionally our customers have been focussing on using our peptides and antibodies for investigating biological systems to identify lead targets for drug discovery but now they also are interested in  monitoring biomarkers so that the developed treatments can be used on the appropriate patients. It is a fascinating area and seems to bring the researchers closer to the patient.

The meeting was held in the the San Diego Convention Center, this center is huge and the exhibition was well run as they specialise in big meetings such as this. The center is right next to downtown San Diego which was very convenient for conference attendees. The city itself is very pleasant and judging by the amount of people around the city with conference badges everyone was enjoying their visit both to the conference and also to the city.

We did suffer from the “June Gloom” which is a coastal fog which blocks the sun in San Diego at this time of year but when the Sun did break through it was very pleasant. Apparently June is renowned as the month of the year not to visit San Diego but we were there for the science and not the sun. Our head chemist Dr Laurent Caron was also in the city at the same time attending the American Peptide Society annual meeting and stayed on for a few days after the conference and apparently it cleared up and was beautifully sunny as soon as the sales team left!

We had interest in custom peptides and antibodies at this meeting so it seems that our services are required within this area of the research world so thanks to those people who suggested we attend.

The meeting next year will be held in Philadelphia from June 8th-12^th.

The first session entitled Peptide Structure and Drug Discovery was a succession of talks from various peptide companies. Sadly, the attention was not always in the detail and some presentations lacked depth. Thankfully, Don Wellings saved the day with some interesting ideas.

Don is a regular speaker at peptide meetings. He is interested in developing cheap technologies for peptide synthesis and is trying to raise interest in his glass beads technology. The idea is to use cheap jewellery beads made on ton scale and coat them with a resin for peptide synthesis.  Another idea is to polymerise oligolysines (made by bacterial fermentation for use as food preservatives) using various cross-linkers.

The second session (Peptide Therapeutics) mainly involved institutional and academic speakers. Mike Gait (Medical Research Council), a world expert in the synthesis of analogues of DNA/RNA such as Peptide Nucleic Acids (PNAs) and Peptide Morpholinos (PMOs) presented some work on the conjugation of analogues to Cell Penetrating Peptides (CPPs). Mike is a very engaging speaker and his presentations are always full of good content. Later, Prof. Rob Lizkamp (Utrecht University) described some dendrimer constructs and some discontinuous epitopes mimicry using his scaffolds. The paper is available online (free of charge!) if you are interested in the triazacyclophane (TAC) scaffold.

Mixed feelings at the end of conference, some good and some bad…

According to OBN around 750 people attended the meeting and it certainly seemed to be busy and vibrant which was good to see. CRB had an exhibition stand at the meeting and took part in the partnering meetings to establish some new business relationships and meet with old acquaintances.

This meeting was very much focussed on investment and the seminars and partnering event reflected this so this was perhaps a bit of an unusual meeting for us but it was encouraging that we still met with actual scientists with an interest in real lab work and peptides. Companies represented at the meeting ranged from traditional big pharma down to 1-man virtual biotech companies so it covered the whole breadth of the industry.

The meeting was potentially more useful from a business development perspective as we are currently seeking new partnerships and looking for interesting technologies. The partnering system was extremely well run and plenty of time was allocated for good conversations to occur to facilitate this. If you have anything you think we might be interested in please do feel free to contact us.

On a lighter note as the event was set at Newbury racecourse we had a better view than most conferences when walking to the partnering hall. However this did give good views of the golfers in the middle of the course having fun whilst the conference attendees were hard at work!

It is of course always enjoyable being away from headquarters and meeting with our customers, suppliers and potential future collaborators. We hope to see many more of you throughout the year.

Iain

The exhibit hall had large booths from pharmaceutical companies (complete with complementary cappuccino) extolling their pipelines of compounds in the clinic.  Exhibit booths from equipment manufacturers, CRO partners, contract employment firms and research suppliers like CRB filled two massive rooms where poster sessions were held.

As exhibitors, time for talks and posters are limited.  We talked with about 200 researchers and had brief visits to our exhibition booth by many more.  Researchers seemed to be predominantly from academic institutions versus pharmaceutical companies; perhaps a sign of declining research budgets in pharma.

Inquiries ranged the gamut from simple to complex peptides, fluorescence, custom antibodies and radiolabelling.  Speaking about post-translational modifications and preparing antibodies to detect these was a popular point of discussion seeing that methylation and phosphorylation events are both important markers in cancer signaling.

This was our third consecutive year exhibiting at AACR and, although attended by fewer researchers, it seemed that interest in us was overall higher than before.  I’m sure we’ll attend again in 2012.

David

Drug discovery researchers are now looking at molecules other than the typical, small molecule effectors we are so familiar with.  The days of the blockbuster pill are numbered:  Binding pockets must be set up perfectly in the receptors and, without very tight interactions with a receptor; high efficacy becomes difficult to achieve.  Doses escalate from a few milligrams to higher amounts leading to off-target effects and toxicity.  Small molecule drugs are excellent in oral dosing, bioavailability and crossing cell membranes and being transported in and out of their environments to do their jobs.  They are inexpensive, have high stability and familiar to all patients.

Biologics such as protein and antibody therapeutics are at the other end of the spectrum.   Large macromolecules can effect protein-protein interaction targets with very large binding and irregular binding surfaces.  They are based on natural ligands and need little structural modification for efficacy.   Dosing and pharmacokinetics and metabolism (where a drug goes and how long it lasts in that condition) are difficult and solved with injectable or highly technical dosing solutions.  Drugs like Enbrel, Herceptin, and Epogen all are examples of antibody therapies made using recombinant DNA technology.   These drugs are somewhat unstable, injectable only and quite expensive therapies used only when other approaches are exhausted.

These two classes are only 10% each of the chemical space which drugs might exist.  The remaining 80% remains between the very small and very large molecules.  As presented by Greg Verdine at the 240^th National American Chemical Society meeting, somehow, this massive area needs to be explored for new therapies in disease cure and prevention[i]

Crossing the bridge between these are peptides.  Peptides are smaller than proteins (although no hard and fast rule exists how much so) and can behave like small molecule drugs.  They have high affinity for their receptor and have been designed so they don’t cross react with other targets.  Their downfall can be absorption (like protein and antibody drugs) and half-life in the body.   Insulin, GLP-1, salmon calcitonin and others have entered the market as viable alternatives in this space[ii].   Yet, like proteins, they need to be injected (or highly formulated), can be cleared and processed quickly and don’t easily cross cell membranes to reach their receptors.

Promise, as reported by Verdine, Sawyer, Walensky and others, comes from stapled peptides[iii].  Regular peptides need to bind to their receptors in exact fashion by adopting a specific structure.  However, they are floppy, random, coiled or unordered and need to overcome this entropy by raising their binding energy to fit the receptor’s requirements.  By “stapling” these, one can force the peptides to adopt specific conformations and overcome that entropy factor allowing tight binding.  Another consequence is that the peptides are less visible to destruction and clearance via natural clearance processes.  They also, in some cases, have the ability to cross cell membranes and be transported to their receptors.   To see the dramatic effect stapling can have, check out this video showing just how much energy savings stapling provides on the peptide and how the binding residues line up for presentation (and the great techno music)[iv].

These rigidified peptides have been explored for decades using different approaches.  Di-sulfide knots are nature’s way of making a peptide tightly folded.[v] I to i+4 cyclic lactams are similar to the stapled peptides, but require complex solid phase chemistry to prepare[vi].  Helix inducing caps have been synthesized to attempt to induce an environment where the αhelix is the strongly preferred conformation.[vii] Stapled peptides give chemists the ability to induce this helical constraint in the last step of synthesis after purification and analysis of a linear material.  This highly selective clipping by either metal catalysis or photoinduction is highly selective, so the chemist has the ability to make well characterized and purified molecules through the whole process[viii].

Successes are starting to roll out.  Aileron Therapeutics is based on this technology[ix] and has 3 demonstrated successes in R&D.  Chemistry different from the original cyclization has opened more routes to helix induced peptides.[x]

Such elegant chemistry requires expert hands.  Experience in peptide and organic chemistry syntheses give higher success rates.  CRB has PhD level organic synthesis and peptide chemists for synthesizing such peptides, as well as any needs for un-natural amino acids, novel amino acids or highly complex structures.  If you’re interested in peptide drugs or peptide projects which could use a burst of advancement, please contact us for more information.

1. http://tinyurl.com/479lvfl
2. http://qjmed.oxfordjournals.org/content/92/1/1.full
3. http://pubs.acs.org/cen/coverstory/86/8622cover.html
4. http://www.youtube.com/watch?v=WScPbvUwDno
5. http://www.ncbi.nlm.nih.gov/pubmed/7576659
6. http://onlinelibrary.wiley.com/doi/10.1111/j.1399-3011.1998.tb00664.x/abstract
7. http://www.ncbi.nlm.nih.gov/pubmed/11603974
8. http://tinyurl.com/4vl5tqo
9. http://www.aileronrx.com/index.php
10. http://www.innovations-report.com/html/reports/life_sciences/techniques_stapling_peptides_spur_development_drugs_169828.html

The first session on Isolation, Characterisation and Production of AMPs was a succession of computer simulations and presentations of new techniques to study the mechanism of action of antimicrobial peptides. One of the highlights of the day was the talk given by Matthew R. Hicks ,  a senior research Fellow at Warwick University, who presented the linear dichroism technique that he has been working on to investigate the orientation of peptides in cell membranes.

A buffet lunch was served in the Analytical Lab (this is the only time you are allowed food in the lab!) and the second session on Therapeutic Applications started with a presentation from Frances Chadbourne (Steve’s Research Associate) about anti-leishmanial activity of some temporin peptides and some awful pictures about the effect of this disease. Eefjan Breukink (Utrecht University) closed the first day as ‘keynote speaker’ with some very interesting ideas about the design of targeted pore-formers to kill bacteria.

The second day was about the Chemical and Biological Approaches to modifying AMPs and seeing all these structures on the white board was just every peptide chemists’ cup of tea. Aletha Tabor from University College London gave a passionate talk about the full synthesis of Nisin (a natural product and food preservative – E234) and more generally lantibiotics. Nisin is a very complex peptide with 5 lanthionine bridges and its total synthesis is of interest to build similar structures.

Overall feeling was very positive. Steve certainly did a very good job putting this meeting together and it was a very friendly gathering. We look forward to the next edition!

Fluorescence in biotechnology continues to be a baseline need for multiple applications.  Following the drug discovery flow process from early target discovery, through compound nomination, toxicology and clinical and post-clinical, the use is always there.  Even those annoying eye drops the ophthalmologist uses to reveal corneal scratches rely on fluorescein, the most basic and common dye used[ii].

In early discovery and target validation, biochemical assays are needed.  Radioactive ligand binding assays still have a place and are very useful, but these have reduced throughput, can have nearest neighbour artifacts and can use ligands which expire after a few half-lives and have costs and regulations associated with their disposal and storage. Fluorescence polarization[iii](FP) has supplanted the protein-ligand binding (be it DNA, RNA, peptide or small molecule) assay and uses dyes of longer wavelength to avoid matrix effects.   FP needs fluorophores which have long lived fluorescence half times, polarize well and have the perfect linkage and spacing to the molecule being studied.  A modular approach such as cysteine peptides and maleimide activated dyes makes finding an ideal ligand less onerous.  Protease assays relying on Fluorescence Resonance Energy Transfer (FRET) or time resolved FRET (trFRET) have become fundamental tools for activity, specificity and kinetics assays of drug targets.   Fluorophores on molecules in near proximity quench each other’s fluorescence and when these dissociate, signal is observed.  Protease cleavage of a dual-tagged peptide substrate, such as the β-secretase cleavage assay is a common variety of this.[iv]

But the technique has applications outside of drug discovery as well.  In-vivo imaging of tumours using fluorescent probes gives a unique, long lasting image of tumor distribution by delivering materials (antibodies, Q-dots, peptides, etc.) to the tumor, binding with and illuminating it.[v] Surgery using “Light-up peptides”, which are fluorescently tagged peptides, to visualize exact locations of nerve structures means less problematic outcomes.[vi]

We at CRB are well versed in fluorescent tagging of peptides and antibodies.  Our licenses with Life Technologies / Molecular Probes allow us to provide custom peptides tagged with Alexa Dyes, BODIPY, Rhodamine and fluorescein derivatives.  Our exclusive license with GE-Amersham allows us to provide custom peptides with Cy dyes (as used in the Ballou paper above).  Our latest partnership with Cyanagen bring the next generation of Chromis dyes into our portfolio providing dyes with optimum brightness, stability, charge flexibility and known published structures.

Please contact us (and check out our dye selector) at our website www.crbdiscovery.com , by email atcrbsales@crbdiscovery.com or at +44 (0)1642 567180 for a consultation about how we can add these reagents to your research.

For further information, CRB regularly publishes a newsletter with links to interesting stories within our field and focussing on our custom service capabilities. Check out the last issue : Fluorescent labelling across the spectrum

1. http://www.youtube.com/watch?v=YvN8zFhWn04&feature=related
2. http://www.nlm.nih.gov/medlineplus/ency/imagepages/9330.htm
3. http://calvino.polito.it/~gasparini/fp_introduction1.pdf for a primer on the physics of the assay
4. http://www.sigmaaldrich.com/life-science/cell-biology/learning-center/bace1-assay-kit.html
5. http://onlinelibrary.wiley.com/doi/10.1021/bp970088t/abstract
6. http://www.nature.com/nbt/journal/vaop/ncurrent/full/nbt.1764.html