Category Archives: Startup

Cannabis sativa

I work on cannabis.

As a PhD biochemist and former cancer researcher, it feels a little dirty to admit. But let me explain…

I love plants. My favorite plant is Sarcodes sanguinea, a native of the high Sierras that is a brilliant red hue because it lost it’s chlorophyll and parasitizes fungi for food:

A fabulous Sarcodes seen while riding my cyclocross bike on the East side of Lake Davis before the Lost and Found Gravel Grinder, June 2017.

I planted lemon, lime, fig, pomegranate, olive, pear, and apple trees in my back yard. I also have a rockin’ garden, and I love teaching my kids where their food comes from:

My main garden in full force, June 2017.

I also have a soft spot for interesting plant chemistries. While at U.C. Davis I worked with antioxidants produced from broccoli and other cruciferous vegetables. I have worked with the biochemical pathways producing the stevia sweetener, and contrast that with working on the genetics of noxious plants that are toxic to humans.

I am an  endurance athlete, and I like to cycle or trail run for hours. Mostly from the health benefits, but I do enjoy the endorphin rush from an epic workout. I’m generally risk-averse, and I’m an Eagle Scout in most senses of the stereotype.

But I work on cannabis. Or more specifically, cannabinoids that originate from the cannabis plant.

Cannabis sativa. Pot. Weed. Marijuana. Ganja. Reefer. (Insert your favorite slang here). Yeah, that cannabis.

It is weird to me how some people think that I am part of a counter-culture movement full of pot-smoking hippies (or whatever the equivalent is for the millenial generation – pot smoking, avocado-toast-eating, millenials?). In actuality, we are a bunch of over-educated, lab-coat clad nerds, drinking coffee and geeking out about the amazing chemistry of cannabinoids and the corresponding human cannabinoid receptor system.

Nerds!!!

As Alexander Shulgin wrote in PiHKAL, “Among the drugs that are currently illegal, I have chosen not to use marijuana, as I feel the light-headed intoxication, and benign alteration of consciousness does not adequately compensate for an uncomfortable feeling that I am wasting time.”

None of us partake in cannabis for recreational or medicinal uses, and whenever a question on human use comes up- we are forced to consult Google to learn about typical usage and exposures. I personally, and our startup are fully compliant with all state and federal regulations, and we have all necessary approvals to perform our research. When it comes down to it, we are just doing science. Some plant biochemistry, some mammalian receptor biology. But our work could have an impact on inflammatory bowel disorders, colon cancer, and some other potential applications that could be pretty damn cool.

A friend from grad school was concerned that by researching cannabis, I may be a marked scientist- unable to rejoin the ranks of the normal scientists hard at work on cancer, neurobiology, and other respectable areas of study. I completely disagree. What I have found is that the cannabinoid and cannabinoid-receptor research field has been stifled by undue regulatory pressure for so many years, so instead of being a dead-end for science, it is ripe for discovery.

Pharmacological uses of non-psychotropic cannabinoids

The cannabinoid receptor is the most abundant G-protein coupled receptor in the brain, hence, people get really high with THC exposure (an agonist). CBD on the other hand is an inverse-agonist, so it has somewhat of an opposite effect on our receptors and doesn’t get people “high”. It is on the fast-track to be approved for treatment of specific epilepsy subtypes, is in multiple clinical studies for schizophrenia, and has tremendous potential as an anti-inflammatory. So the pharmaceutical applications are real, and have even been vetted by the Food and Drug Administration.

Cannabinoid receptors are also found throughout the body, and to use the “lock and key” analogy for enzymes- we have discovered locked doors throughout the body- now we just need to figure out the keys, and what doors we can open with this new knowledge. The therapeutic  potential for cannabinoids is unmistakable. I look forward to seeing what doors we can open with our work.

-BZ

Startup Quotations

A fun image borrowed from yhponline.com on their UK Startups page

As I approach my fourth year of grinding it out in a startup biotech, I thought I would collect my favorite quotations of startups and productivity. As I read more about business and startups I keep finding new quotes that encapsulate the feeling of a startup, so I hope to add to this list as time goes on.

Move fast and break things. (attributed to Facebook, and humorously depicted on XKCD).

A ‘startup’ is a company confused about: 1. what its product is, 2. who its customers are, 3. how to make money.” – Dave McClure of 500 Startups.

The reality is, Silicon Valley capitalism is very simple:
Investors are people with more money than time.
Employees are people with more time than money.
Entrepreneurs are the seductive go-between.
Marketing is like sex: only losers pay for it.” – Antonio Garcia Martinez in Chaos Monkeys: Obscene Fortune and Random Failure in Silicon Valley.

Done is better than perfect. (described as a Facebook slogan in Chaos Monkeys).

A good idea on Monday is better than a great idea on Friday. (not sure where I heard this one).

Build. Measure. Learn. (repeat).  – Eric Ries, The Lean Startup

Startups are an odd creature that I am still working on taming (assuming you can tame it!). The energy and speed are addictive, and despite all of the struggles, I wouldn’t trade it for anything.

-BZ

The Budget Biotech

The bench beaver is always watching you

I have been toying with the idea of starting my own biotech startup in my garage, and want to determine the lowest amount of upfront investment it would take to set up a functional molecular biology lab.

OK, a little background to help the discussion. Straight out of grad school I took a job with a startup biotech developing strains for fermentation of high value small molecules. When I arrived the lab had a few tables and some basic equipment, but being that an electrical engineer did the initial purchasing it wasn’t quite ready for much biochemistry. So my first job was to outfit the lab with the necessary equipment and consumables to get things moving. I quickly realized how expensive laboratory equipment and research reagents were without the generous academic discounts I had grown accustomed to. We pieced together a mixture of new and used equipment, took advantage of lab startup promotional deals, and ultimately assembled a fully functional molecular biology lab on a pretty modest budget.

Taking what I learned from setting up my first biotech lab, I know I can do better. How much better? I think it should be possible to assemble a functional molecular biology lab for under $1,000. But wait you say- that is just enough to buy a decent set of pipetters- how could you set up an entire lab for that?

Precisely. That is the heart of the challenge. To create a budget-minded biotech that is functional, but doesn’t cost an arm and a leg. As I wrote up in a previous article, “biotech shouldn’t be so dang expensive!”

I think it all depends on how you define the core functionality. To me, the minimum functionality needed is to clone genes and express proteins. I am focusing on the steps from source DNA to expressed recombinant protein. So basically from PCR through cloning in E.coli, to expression of proteins.

Things I am not including in the $1,000: Analytical (in my case HPLC or LCMS), basic infrastructure like tables, stools, computer, etc., and little things like surge protectors or extension cords. And I may take the liberty of culturing my own competent cells to cut down on that cost. Vector backbone will be an open-source design to get around IP conflicts and keep things cheap. We’ll see how far we can stretch $1,000 and go from there.

I am looking mainly at functional but well-loved equipment on ebay and craigslist, and even some homemade or hacked projects capable of carrying out the job at hand. EBay has some amazing deals. So does Amazon (I usually avoid Thermo subsidiaries as much as I avoid Wally World!). And if you are willing to do some soldering, coding, and 3D printing, you can make just about anything in the lab. I acknowledge that reliability will undoubtedly be less than brand new equipment still on warranty, but of the three-legged stool of quality, cost, and time- this exercise is focused primarily on cost.

Here is my initial list of equipment, projected cost, and notes on procurement. Let me know what you think, where you think I can cut the cost down, or if there are any glaring errors in my choices or logic. I plan to revise and post updates to my budget biotech quest, so hopefully this is only the beginning!

-BZ

EQUIPMENT
Item Cost Notes
pcr – thermal cycler $200.00 used… Ebay. Or ghetto fab water bath and servo setup?
gel rig $53.00 kit from iorodeo.com
light box $80.00 mini kit from iorodeo.com or build myself
pipetters $100.00 2 pipettes at 50 each. P20 and P200 to start
water baths $12.99 crock pots with arduino thermostat (DS18B20 and SSR)
-20C freezer / fridge $50.00 total budget – cheap chest – or used on CL
incubator $50.00 Cooler/chest freezer with hairdryer/heatgun heater and extra fan? Or foam insulation box.
autoclave $83.99 pressure cooker- presto on amazon. Ikea?
centrifuge $80.00 used Brinkman on Ebay
flame for plate work $60.00 Ebay flame/regulator kit
Micro scale $25.00 amazon
microwave? $20.00 craigslist
gel power supply $15.00 homebuilt (budget molecular biology power supply)
rotary shaker $140.00 used old NewBrunswick on ebay
pH meter $20.00 Amazon or Ebay
TOTAL EQUIPMENT: $989.98
CONSUMABLES
Item Cost Notes
tips $10.00 Ebay- bulk tips – need upfront tip box
weigh boats $10.00 or papers
DNA stain $68.00 SYBR Safe – cheaper alternatives with blue LED illuminator?
gel buffer TAE homemade
DNA polymerase $39.00 mastermind from geneandcell.com
dNTPs included in mastermind mix
DNA buffer TAE homemade
gel extraction kit NA45 paper? Simple protocol? Kit…
1kb ladder $89.00 geneandcell.com
load dye bromophenol blue
comp cells homemade – but lack of -80 will affect competency
Total Consumables: $216.00
Total Combined: $1,205.98 over budget already! And chemicals not even included yet!

Budget Molecular Biology Power Supply

For my first blog post I have chosen to write up a project I did very early in my startup days. The theme of this post is that biotech shouldn’t be so dang expensive.

While setting up a molecular biology lab for a startup biotech, I was shocked at the industry prices on equipment and reagents, something I took for granted while still in academia. Nowadays I am better at vetting Ebay sales for used equipment, but at the time I thought there were two options: new or homebrewed. So I made a simple 120v DC power supply to run SDS-PAGE gels for about $20 in parts. I think you could probably get the parts down to about $10 per power supply.

Disclaimer: I am a biochemist, and not an electrical engineer or someone who designs electronics for a living. Electricity hurts. This post is meant to be informative, but not fully instructional. Use this information with extreme caution, and don’t blame me if you shock yourself or catch something on fire!

So it all started when I needed another power supply to run my SDS-PAGE protein gel system. I didn’t want to buy an expensive power supply because I just couldn’t justify spending hundreds of dollars for something I wasn’t going to use much. So in the early days I used alligator clips to tap into the leads from my BulldogBio DNA agarose gel box. This was pretty sketchy, so after a few runs I decided to open up the power supply and see what was inside. I saw only two switches, a fuse, and an electrical component that was new to me. After googling the writing on the module I learned that it was a Bridge Rectifier- basically a big rectifying diode setup that can convert the AC into a choppy DC supply. The Bulldog power supply obviously worked well for running gels, so I didn’t think that the rough DC signal was a problem. Inside the BulldogBio unit there was also a switch to switch the negative/ground to the electrophoresis rig between the true AC ground and the (-) leg of the bridge rectifier (I think this is how it switches from +60vDC to 120vDC), but I didn’t implement that in my power supply (yet). So I pieced together the parts, built the power supply and it worked!

IMAG1280
Link to fullsize image
IMAG1279
Link to fullsize image

Bill of materials:
Electronics enclosure of your choosing
120vAC Power Inlet (and a cord- or just run the cord into the box)
Double Pole Single Throw (DPDT) switch rated to >5A AC (I used a DPDT switch I had laying around)
Fuse holder and quick burn 1A fuse (you can get one built into the power inlet also)
Bridge Rectifier (I bought an overpriced one from Fry’s)
Banana Plug Terminal
A scrap piece of proto board that I had to drill out to fit the big terminals of the bridge rectifier
Some scrap 20ga wire and blade fittings
Solder and iron to connect to the fuse holder and power terminals
Heat shrink tubing to cover the naked power terminals (I didn’t, but I would next time)

Notes:
I definitely would not use this for sensitive electronic equipment, as the DC output is probably pretty rough. It would require some beefy capacitors and voltage regulators to fine tune the output, but that is way beyond my knowledge. The ability to switch between 60vDC and 120vDC would add on a couple of dollars, and would give me the ability to run a gel a little slower. This would likely only require a single pole-double throw switch to switch the gel negative pole between the true AC ground and the (-) leg of the bridge rectifier. But I haven’t gone that far, for what it’s worth.  Also, I haven’t really tested how much heat is generated by the bridge rectifier after an hour of use. The overall box stays cool, and all components look fine (read: no meltdowns yet). If you are running multiple gels or need to push more current through the rectifier, consider getting one with a heatsink, or at least use a fan to move some air through the box. Also, I haven’t really tested how much heat is generated by the bridge rectifier after an hour of use. The overall box stays cool, and all components look fine (read: no meltdowns yet). If you are running multiple gels or need to push a higher amperage, then definitely keep an eye on the box and component temperatures.

I’ll try to sketch out a schematic when I have a moment- or maybe a Fritzing diagram if I’m feeling saucy.

-BZ