News on trends and issues in the biopharm and pharmaceutical industry. Commentary on current events,clinical pipelines, facility expansions, competition, technology, legal and economic matters. M&A and licensing activity across the globe.
I am a Director of Research and Devopment for a leading supplier to biopharmaceutical producers. The views expressed are mine. I do not speak for any company or corporation.
The Big Red Biotech Blog is happy to recognize its 25,000th page view this morning! It's a beautiful Sunday morning here in Saint Louis and nothing goes better with Sunday Brunch than popping open some champagne.
So here's to my readership! Celebrate with me. Not only do we hit another pageview milestone but the Rangers and Giants pull upsets to reach World Series. And Mizzou wallops OU for a change. It's a beautiful day in the neighborhood.
Typical. Late again with this post. This recipe is adapted from the Creole Gumbo and All That Jazz cookbook by Howard Mitchum.
This is Baked Flounder a la Creole II (page 190). I've subsituted flounder for Red Snapper.
3-4 8-oz flounder fillets
1 16-oz can of tomatoes
1 6-oz can of tomato paste
2 scallions with 3" of their green leaves, chopped
1 medium onion, chopped
1/2 green pepper, chopped
1 clove garlic, minced
2 tbsp fresh chopped parsley
1 rib celery, chopped
2 bay leaves
2 lemon slices
1 tsp chili powder
1 stick butter
2 tbsp flour
Salt and freshly ground black pepper to taste
Here we go, let's start with the sauce. Melt butter in deep pan, add the flour and stir until blended. Add the tomatoes, tomato paste, scallions, onion, green pepper, garlic, parsley, and celery. Add the bay leaves, lemon slices, chili powder, salt, and black pepper. Basically, add all the damn ingredients to the butter/flour roux. Cook and stir until vegetables are soft - if you can see them through the tomato ingredients - and translucent. Add enough hot water to make the sauce semi-fluid, and let it boil gently for 20 minutes. Remove the bay leaves and lemon slices.
In the meantime, preheat the oven to 350 F. Place the fish fillets in a greased oven pan, cover with the sauce (you can reserve extra sauce), and bake for 20-25 minutes or until the fish flakes easily when tested with a fork. Lift the fish out with a spatula (treat gently!) and place on preheated (or not) serving plates and cover with sauce. Decorate with lemon slices, sliced olives and parsley sprigs. Serve at once.
I accompanied dish with brown rice, broccoli florets, and fresh baked bread sticks. Serve with hearty red wine - I paired with Dona Paula Estate Malbec, Mendoza, Argentina.
This may be a bit mean, but I couldn't resist the temptation. Fierce Biotech published its second annual Biotech Graveyard list. The bad news, 8 companies went out of business without a buyer or money to forge onward. The good news, that represents a 50% decrease from the 16 companies who went under in 2009. Here is the 2010 Biotech Graveyard:
Anesiva - South San Francisco - Feels no pain
AutoImmune - Pasadena, CA - Self-liquidating!
ConjuChem - Montreal, Canada - Au revoir mon ami!
Hawaii Biotech - Alea, HI - Aloha
Middlebrook Pharmaceuticals - Westlake, TX - No more middleman
Neuropharm - United Kingdom - Keep stiff upper lip, Chaps!
Vio Pharmaceuticals - New Haven, CT - Shoulda' had more insurance
VitalMedix - Hudson, WI - Vital signs are very weak!
More self-congratulatory ecstasy, I've received my 20,000th page view today -- since I began this blog on January 1st. My annualized run rate is now up to 55,000 page views - yeah! While I know for certain there are bloggers out there, who greatly exceed this rate of traffic -- allow me to pat myself on the back. Ouch! I wrenched my arm again.
Don't burst my bubble for now. And for those of you who contributed views, thanks!
OK. I'm calendar challenged and I admit it. That's the first step toward a cure. It's a somewhat overcast Saturday in St. Louis and the Cards lost again. Not much going on - so I decided to retreat to the kitchen this afternoon while simultaneously watching Michigan-Notre Dame - which was a good game.
I made two variations of the same recipe out of the Pink Adobe Cookbook by Rosalea Murphy, owner of the restaurant of the same name in Santa Fe. Today's dishes were Poulet Marengo and Cotelletes de Porc Napolean.
Roselea explains in her cook book that Poulet Marengo was a tribute to Napoleon's victory over the Austrians at Marengo in 1800 concocted by Napoleon's personal chef, Dunand, to celebrate the victory. As with all things Pink Adobe, this recipe has Rosalea's flair added with modern ingredients.
Don't worry. It's easy or I wouldn't be able to attempt it.
1 cup flour plus another 2 tablespoons
1 teaspoon salt
1 teaspoon paprika
1 1/2-2 lbs boneless chicken breasts
2 tablespoons butter
2 tablespoons olive oil
1 cup Madeira
1 clove garlic, chopped fine
4-5 large mushrooms, sliced
12 pitted black olives (or more)
1 small bag pearl onions (you figure it out)
1 16-oz can of tomatoes
1 4-oz can of tomato juice
1 bay leaf
1 cup chicken broth
Cotelletes de Porc Napolean
4-6 center cut loin pork chops, 1 inch thick
OK. The meat portions that I listed above have all been cut in half because I wanted to make both chicken and pork from the same recipe. All other ingredients under Poulet are intact and I split them between the two meats in separate baking dishes. Capice.
Preheat oven to 325 F. Put 1 cup flour, plus salt and paprika into a coating bag. Add one or two pieces of meat at a time and shake until coated. In skillet, heat butter and olive oil. Brown chicken and pork, one or two pieces at a time. You do not have to cook through. Pork can be browned less than chicken. Add more oil as needed, but the meat should not be greasy.
Place browned chicken or pork fillets in baking dish. Cover with 1/2 portions of ingredients above. Add Madeira wine. Add garlic on each piece. Then add olives, mushrooms, onions and tomatoes. Last add the tomato juice and a bay leaf.
Cover baking dishes and bake for 2 hours. At end of cooking time, remove chicken and pork to serving platter and keep warm. Meat will be fork tender so treat it gently.
With wire whisk, mix remaining 2 tablespoons of flour with the 1 cup of chicken broth. Combine the juice from baking dishes and bring to a boil. Beat in broth until it thickens. If it becomes too thick, then add some Madeira to thin.
Plate chicken or pork. Serve sauce separately. This meal goes well with green noodles or other colorful pasta. Serve with Chassagne Montrachet (or whatever wine you prefer). Bon Appetit!
For this lesson, Scott refers us to the Dr. Seuss parable about the Sneetches - some with and some without "stars upon thars". Many companies make the mistake of confering innovation groups with "stars upon thars" which naturally leads to resentment from those without stars. Unfortunately, those without stars reside in the base business and must play a key support role if the innovation group is to succeed.
If you create a sense of have and have not with regard to the confirmation of "stars upon thars" for some, you make a big mistake at the beginning that will likely doom the innovation effort.
I'm feeling less depressed and anxious already, Man!
An old idea is resurfacing as new. Dr. Franz Vollenweider, Zurich's University Hospital is advocating the use of psychedelics like LSD, ketamine and psilocybin (Magic Mushrooms) for the treatment of depression, repressed memories or bipolar disease in some patients.
Wow. I really fell down on the job with my recipe posts - missing June and July completely. Ooops! I did make something this weekend though that I enjoyed quite a bit and thought I would share. These two recipes come from the Pink Adobe cookbook by Rosalea Murphy. The Pink Adobe is a famous restaurant in Santa Fe -- one that I highly recommend that you visit if you are ever n that beautiful community. It's an institution and well worth the visit.
Without further fanfare, here are Lasagne Monte and Many Bean Salad.
1 lb lasagna noodles
2 lbs boneless, skinless chicken breasts, poached
3 cups fresh green chiles roasted or 1 27-oz can of chiles (I used chopped & worked fine).
1 1/2 cups ricotta cheese
1 1/2 cups sour cream
1 8-oz package of sliced mozarella cheese
1/2 cup of sliced black olives
To poach chicken, place in 4 qt sauce plan with 3 cups of water. Bring to boil. Add 1 carrot, 1 celery stalk, 1/4 onion, salt & pepper. Bring to boil and reduce heat to simmer for 45 minutes. Let cool in chicken broth. Broth may be strained and retained (frozen) to serve as chicken stock in future.
Cook noodles according to package instructions. In 9 x 15 dish, place first layer of noodles. Slice poached chicken into strips. Cover noodles with 1/2 the chicken. Cover chicken with 1/2 the green chiles. Mix ricotta cheese and sour cream. Cover chicken and chiles with 1/2 the cheese mixture. Cover with next layer of noodles and repeat the process. Place final layer of noodles on top. Cover top layer with the sliced mozarella cheese and then the chopped black olives. Pre-heat oven to 325 F and bake, uncovered for 30-40 minutes until heated through. I made dish the night before and refrigerated before baking which seemed to work fine.
Many Bean Salad
2 tablespoons mild cider vinegar
1/2 teaspoon sugar
1/4 teaspoon salt
1/8 teaspoon freshly ground black pepper
1/2 teaspoon Dijon mustard
5 tablespoons olive oil
1 cup each canned, drained beans - I used kidney, black, pinto, navy, garbanzo & green beans. The mix of colors is impressive.
1/4 cup chopped green pepper
1/4 cup chopped red pepper (my addition)
1 cup chopped red onion
1 medium tomato chopped
1/2 cup of mayonnaise (I omitted)
Prepare dressing by mixing all ingredients except the oil. Mix in olive oil and shake well (can use screw top jar). In large bowl, combine all the salad ingredients - except tomatoes & mayonnaise. Use rubber spatula to mix to protect integrity of beans. Pour dressing over salad and mix gently. Refrigerate if desired. Just before serving mix in tomatoes & mayonnaise.
I served the lasagna and salad with warm crusty bread. I would normally drink with a hearty red wine, but chose to have an amber pale ale instead - which I enjoyed. Bon Apetit!
Derek Lowe writes the very very good and very highly read In the Pipeline blog. Derek has a way of getting right to the point with a nice blend of bluntness and sardonic humor without being mean spirited. Sometimes his posts just make me smile. Here's one such post from today on the topic of Open Source discovery.
I'm actually a backer of the concept and think that companies really should try to explore its potential -- by testing and operationalizing it. However, Derek makes a good point in his blog. Too often proponents on the topic - provide a lot of talk but don't seem to be able to deliver the concrete actions or proof of concepts.
Maybe this post hit home a bit more as I received the following note from a colleague commenting on a current discussion on "open innovation" in one of the innovation groups that I participate in. My colleague wrote:
To me it's unbelievable how little capability to analyze things have most of the people who are posting comments on that discussion group!!! They got "stuck" into that Soft skills topics they can read in books of Change management, Leadership, etc, but when it comes to Hard skills and concrete innovation they do bla-bla-bla...
Presumably it is hard to articulate the "hard innovation ideas" as they are hidden in the ball of fuzz that Derek refers to in his post. Maybe we need to contact Rush to help us find the answer as he seems to know a lot about Fuzzballs.
In this investigation, it was determined that PVA was causing the filtration issue and that the problem was related to variation in the degree of hydrolysis seen with the raw PVA.
Subsequent discussions (our data in hand) with the manufacturer eventually convinced them there was a change in the product performance resulting from the hydrolysis effect. It was then determined that an equipment change might be related to this observation.
The manufacturer went back to the old equipment and produced a lot of PVA for SAFC's Raw Material Detectives to test. Lo and behold! The material resulting from the old equipment was more soluble and showed much better filterability. We now have agreements in place to secure only material produced in this manner & thus can make media again with PVA that filters well.
The USA will play Ghana today in a knock-out match with hopes of reaching the Quarter finals in World Cup play. The game starts at 1:30 pm Central time (where I live) from Royal Bafokeng Stadium in Rustenburg. The US side has fared well here in previous matches vs Egypt and the UK.
Good luck to the Team this afternoon. I hope we can cope with Ghana's quickness and no doubt will be battling strong home town African support. This is our chance to reverse the outcome of the 2006 game vs Ghana.
US fans are stoked. Football (soccer) is top of mind right now for many and networks are expecting to top the record 13.7 M viewers set during 1994 loss to Brasil.
I like articles on Innovation. This is a good one by Scott Anthony in the HBR blog. I pass it on as a cautionary tale that really can be applied to any big successful company in any industry. Those of us who have spent careeers in large organizations can recognize a few of these observations and behaviors - I hazard to submit.
This might be a good time (if not already too late) for executive leadership teams throughout pharma to ponder some of these weighty questions.
It was now an overcast day as we return to our story.
What next? The RMDs reviewed all the lot numbers of the PVA from above average, average and below average filtering lots of customer media at large scale. This review indicated a temporal correlation between good and poor filtering lots of PVA used in the media. It became apparent that lots of PVA manufactured prior to Feb 2007 on average filtered better than subsequent lots of material.
The Raw Material Detectives decided to look at selected PVA lots along the time continuum, with known good or poor filterability, in a turbidity assay. In this assay, 6 grams of PVA were dissolved in 100 ml of water with stirring for 60 minutes. Turbidity was measured using a Hach 2100N Turbidimeter and reported in NTU.
Data for the turbidity assay appears in the Table 1 below. PVA lots are arranged chronologically by date of manufacture. Results show a strong correlation between turbidity and filterability. Lots with turbidity below 10 NTU filter above average. Lots with turbidity measurments greater than 20 NTU filter below average.
In the Table the lot numbers have been altered to protect the innocent. The altered lot numbers depict month and year of manufacture. Note, two lots were used in multiple batches of medium and in some cases exhibited above average filtering characteristics - even with the high turbidity number.
Lot 02-2008 (2)
Below & Above Avg
Below & Above Avg
The RMDs had some evidence to support their hypothesis that a change occured in the manufacturing process after Feb 2007 but wanted more.
The PVA lots hydrated at 6% for the turbidity assay were placed in a centrifuge at 3200 rpm. The pellet was dried and weighed. The percentage of PVA in the pellet was calculated. The correlation was high between PVA in the pellet and the turbidity values. Data from 3 of the above lots is shown below in Table 2.
% PVA in Pellet
Enough already! Haven't we shown that the PVA is the issue? It wasn't enough for the Raw Material Detectives. They needed to know why?
Hydrolysis & MW Determination
It was time to analyze the insoluble material in the pellets. The pellets and the supernatants from the above experiment were sent to Chemir Analytical Services for NMR, DSC and GPC (see previous post for more info). The data is summarized below in Table 3.
*Two different batches of lot 01-2009 were sent to Chemir for analysis
The % PVA hydrolysis seen in the pellets was higher than seen in the supernatants. All of the % hydrolysis figures for both the pellets and the supernatants fell outside the specification range of 87-89%. But, the mix apparently met the release specification.
The MW's in the supernatants were lower with a primary peak around 11,000 Da. The pellets were of much higher MW with a primary peak around 21,000 Da. The melting point data did not really show a difference except with one of the pellets in the first 01-2009 lot. We appear to have two subpopulations of PVA.
Smoking Gun Test
The Raw Material Detectives hypothesize that the increased insoluble material found in lots post Feb 2007 resulted in lower filterability -- and set out to reproduce it.
The RMD compared customer CHO media prepared with PVA as normal with media prepared with PVA after the insoluble material had first been removed. Data appears below in Table 4 for the 3 lots previously described.
PVA Treated Supernatant
Thus removal of the insoluble material from the raw PVA resulted in higher filterability. The insoluble material is a subpopulation within the PVA with a higher MW and a greater percentage of hydrolysis. The Raw Material Detectives have identified this as the root cause of the filtration problem.
The Smoking Gun test confirms (as if you are a scientist - you perhaps prefer application of Koch's Postulate).
Acknowledgement: Raw Material Detectives Mike Wathen (old guy pictured center), Steve Wilson, Ben Cutak, Jodi Zobrist (pictured front with mustache) of SAFC Cell Science & Development and Analytical Research & Development groups.
It was a dark and gloomy night (just like last time).
This time two customers contacted the Global Technical Service (GTS) group with complaints of poor filterability of their lastest batches of a custom serum-free CHO medium, being used in large scale production of a commercial biopharmaceutical. This formulation contained about 70 components including soy hydrolysate. We put in a call to the Raw Material Detectives (RMD).
A review of the raw material lot numbers, used in the medium, by the GTS staff identified an association with a specific lot number of polyvinyl alcohol (PVA) CAS 9002-89-5 and poor filtering lots of medium. Thus a clue! The Raw Material Detectives started with the PVA vendor.
Editor's Note:PVA is used primarily in industries like food, paper, adhesives and textiles. Cell culture is a very minor use application by comparison -- i.e. and therefore has reduced influence on the manufacturer/ supplier. PVA is produced by the polymerization of vinyl acetate to produce polyvinyl acetate. This is then hydrolyzed with sodium hydroxide to yield PVA. Possible impurities in PVA would include sodium acetate, methanol, and methyl acetate. PVA should be readily soluble in water but requires heating. Degreee of hydrolysis , molecular weight, particle size distribution, and particle crystalline structure all affect solubility. Optimal solubility occurs at 87-89% hydrolysis, a fine particle size (< 80 mesh), 3.5-4.5 cps viscosity with a 4% solution at 20 C.
Vendor PVA lots were evaluated for D90 particle size, viscosity of 4% solution, and percent hydrolysis of PVA to look for correlation with customer poor filterability results. There was no obvious correlation.
SAFC worked with its customers to develop a scaled down Vmax assay that correlated with the filter difficulties seen at large scale. The following reference was developed.
Vmax <1000 L/m2 were correlated with below average filterability
Vmax >2000 L/m2 were correlated with above average filterability
Vmax 1000-2000 L/m2 generally were above average but not 100%
Based on the above assay, it was decided to to analytically characterize multiple PVA lots with different filterability results in an attempt to explain the poor filtration characteristics.
PVA powders associated with know above and below average filtering lots at large scale were sent to Chemir Analytical Services (Maryland Heights, MO) for the following tests:
Gel permeation chromatography - compares MW properties
Four lots were sent for analysis from the current vendor - 4 above average filtering lots and 4 below average filtering lots. Additionally, the Raw Material Detectives came up with 2 addtional lots from 2 addtional vendors to also test.
Pretty slick set of analytical assays - huh? Unfortunately, there were no significant differences in the physical properties of the various PVA lots tested that were identified by any of these seven assays. WOW! Time to quit? Never. The Raw Material Detectives always get their Man (so to speak). Read the next post for the Rest of the Story.
Be on the alert for the next installment of the Raw Material Detectives this month! Our detectives continue their work unraveling mysteries with raw materials used in cell culture. It's part of their ongoing mission to better characterize and control critical raw materials used in biopharmaceutical processes.
I saw this article in Future of BioPharma (PharmaTimes). Sandoz' Omnitrope is the 1st biosimilar to be recommended for use by the National Institute for Health and Clinical Excellence on the National Health Service.
Omnitrope is a follow on biologic to Pfizer's Genotropin (somatotropin) and the first biosimilar to gain approval in Europe to treat growth problems in children.
My daughter eats a lot of vegetarian items and many of these in some way, shape or form involve the use of squash - usually with fine results. With that in mind, I present Butternut Squash Quinoa Cranberry Salad.
1 cup of quinoa
2 cups vegetable broth
1 medium butternut squash peeled, diced in 1/4 in cubes
1/2 medium red onion diced
1/2 cup dried cranberries
1 sprig rosemary
2 tbsp olive oil
1 tbsp butter
1/2 tsp sea salt
1 tbsp cider vinegar
In sauce pan, bring quinoa and vegetable broth to a boil and then simmer for 25 min. Remove from heat.
Preheat oven to 425 degrees. Add all other ingredients except the vinegar to a baking dish. Coat ingredients with olive oil, sea salt and butter. Roast for 20 min. until the squash is soft.
Remove from oven and add vinegar. Serve baked squash mixture over quinoa. Eat with mixed greens on side.
I'd like to pass on kudos to several of my colleagues at SAFC Biosciences who have volunteeered to participate in the Shawnee Mission West Signature Progams in Biotechnology. In particular, I need to do a shout out for Lynn Davis - one of our Senior Scientists with an extensive background in cell engineering and mammalian cell culture at bioreactor scale.
Lynn has been a lead participant in the program since September 2008. The program is geared toward high school students from grade 10-12 and is designed to expose/begin early preparation for students to career options in biotechnology. Students learn not only about career options, but educational requirements, current trends, and legal responsibilities (is this available for some of our industry CEOs?) through research and guest speakers (like Lynn).
Students learn hands on research skills and techniques through doing an original research project and by onsite experiences at area biotechnology companies. At SAFC for example, we allow students to "job shadow" over the course of four 2hr sessions in a calendar year. The students can learn to make media and aseptic technique, how to passage cells, and are afforded the opportunity to tour our large scale continuous milling manufacturing area.
Other students have learned how to do PCR reactions, assemble bioreactors, sample bioreactors, and put that aseptic technique to the the test. Our current students are bringing cells out of thaw, passaging cells and plotting growth curves. Soon they will be learning to measure IgG levels produced from these cultures on a Fortebio instrument. These students have also toured the QC labs in addition to the milling manfacturing areas.
Students in their own research are doing things like extracting DNA from various cereal grains and examining their GMO status. The program has been very successful. It has grown from 11 students in its inception to 30 for next year.
We're proud to have someone like Lynn on our staff - as well as other team members including Chad Stewart, Brenda Hogan, Barb Martinez, Sandy McNorton and a number of our Cell Sciences and Development group, Rebecca Ball and Katie Hernandez.
It's a pleasure having colleagues who make it their business to give back to education and through whose efforts we are able to provide some "real world" experience and perspective to students who are interested in pursuing careers in biotechnology. It is a great program and it benefits our Staff just as much as the Students who participate - and we're glad we do!
I will soon begin a new series called "The Raw Material Detectives" that will describe unusual cases of raw material misbehavior and how the Raw Material Detectives (our heroes) investigated and resolved the issues. This will hopefully reinforce the need to have good raw material characterization and traceability on products that you are using in the manufacture of your biological drugs.
Now let's give it up for Elvis Costello and the Attractions singing "Watching the Detectives"