A431 cells, treated with predetermined IC50
concentration of novel anticancer agents, fluoresce green and orange-red with MitoPT JC-1. Data courtesy of Zayas/ Carro, Universidad Metropolitana.

Anticancer Effects of Flavonoid Derivatives Isolated from Millettia
reticulata Benth in SK-Hep-1 Human Hepatocellular Carcinoma Cells.
SC Fang, CL Hsu, HT Lin, GC Yen. J. Agric. Food Chem., Jan 2010, 58
(2), pp 814–820.

Mechanisms of Apoptotic Effects Induced by Resveratrol,
Dibenzoylmethane, and Their Analogues on Human Lung Carcinoma Cells.
CJ Weng, YT Yang, CT Ho, GC Yen. J. Agric. Food Chem., Jun 2009; 57
(12), pp 5235–5243.

Gary Johnson

1994-present-President, ICT

1993-1996-Conjugation Chemist, R&D Systems

19989-1993-Supervisor Protein Conjugation & ELISA Development Group, Solvay Animal Health

1986-1989-Immunologists, Biosciences Lab, 3M

1976-1986-Various Lab, U of MN

Gary’s Conatct Info:

• gary@immunochemistry.com
• 952-888-8788  

Inventing Better Ways to Measure Apoptosis 

This profile features another Scientist Entrepreneur. Dr Gary Johnson is the Founder and President of Immunochemistry Technologies LLC (ICT). His company manufactures kits that have the capabilities to quantitatively measure apoptosis effects. This is important to Neuromics, because these are core to many diseases of research interest to our customers. These range from Cancer where apoptosis detection can be used to to visualize the efficacy of tumor killing therapies to Neuroscience where apoptosis could be a root cause of many cognitive and neuro-muscular diseases.

I am excited about featuring Gary. I have been working with him and his team over the past 5 years. They have actively supported my company in providing Apoptosis Research Kits. The strength in our relationship is built on his company supplying best of breed reagents. The feedback I receive from users is overwhelmingly positive. In addition to these kits, ICT is also recoginized for their rock solid ELISA Buffers and Diluents.

It takes a unique blend of business and scientific acumen to build a company like ICT. So let’s start with Gary’s background and experience and then on to the specifics on his company and products and what sets ICT apart from competitors.

Gary’s Background

Gary’s began his career at the University of Minnesota in 1978 where he worked in a variety of labs. There he gained a wealth of experience and expertise in research techniqes. These included chromatography, immunoelectrophoresis, radiolabeling, mass spectrometry,  proton NMR spectroscopy and western blotting.

He leveraged his abilities and became more deeply involved in immunobiology. He  joined Dr. Harry Orr’s lab in 1981. There he used recombinant DNA techniques to study the class I genes of the major histocompatibility complex and he also supervised the tissue culture work. This provided the stepping stone to Dr. David Klein’s lab in 1984. There he studied the difference between diabetic and non-diabetic glomerular basement membrane proteoglycans in kidney disease. In order to do this research Gary developed in vivo or in vivo labeling techniques.

Gary then moved from University to commercial labs. We will see how his growing expertise morphed into the founding of ICT and why his broad knowledge and experise enabled a successful launch of the company.

From 1986 until founding ICT Gary worked at 3M, Solvay Animal Health and R&D Systems. Over his tenure, he worked as an Immunologist, Supervised an ELISA and Protein Purification and was a Conjugation Chemist. Having mastered a unique range of basic and commercial bio-research techniques, the evolution to Scientist-Entreprenuer was a natural next step.

In 1994, Dr. Brain Lee and Gary launched ICT. The company’s early success was in contract assay development. The revenue generated from these programs, has enabled ICT to manufacture and release a growing catalog of Apoptosis Detection Kits.

ICT’s Products and Capabilties

ICT’s provides proprietary probes for measuring apoptosis in vitro and in vivo. These probes are used by researchers  to detect caspases, cathepsins, serine proteases, cholinesterase enzymes, and assess mitochondrial health.Applications include: assessing the efficacy of chemotherapy, to quantifying  neurodegeneration, and early detectionof eye disease, to name a few.

Specific Products Include:

• FLIVO™ Polycaspase Live!, in vivo Apoptosis Kits
• FLICA™ in vitro Caspase Kits
  • Fast!-Use Caspase kits to quantitate apoptosis via active caspases in whole, living cells. These kits do not use ELISA or any antibodies for detection
• FLISP™ Serine Protease Detection Kits
  • Measures chymotrypsin-like protease
    activation in whole living cells.
• Magic Red™ Real Time! Kits
  • Measures apoptosis in whole living, intact cells – no lysis required
• MitoPT™ Kits
• Quantitate mitochondrial functionality and apoptosis

Images: Normal (left) and keratoconus (right) corneal fibroblasts were labeled with Caspase 3 & 7 Assay Kit, green.

Pacing the Field

ICT is setting the pace in Apoptosis Detection by  recognizing and resolving issues inherent in competitive offerings. These include:

1. Difficulty permeating cells.
2. High background problems.
3. Does not bind to early stage apoptotic cells.
4. Not as sensitive as a cell permeant inhibitor probe.
5. Does not bind to all apoptotic tumor cells (Dicker, Cancer Biol. Ther., 2005. 9:1014-1017).
6. Binds positively to normal and healthy bone marrow derived cells (Dillon, J. of Immunol., 2001. 166:58-71).
7. Many in vitro protocols involve lysing the red blood cells before running flow cytometry, this method results in the binding of Annexin V to all of the cells in the sample (Tait, Blood, Cells, Molecules, and Diseases., 1999. 25:271-278).  The inversion of PS and cells containing large amounts of PS may not be related to apoptosis and this adds to the background issues.
8. Does not measure a process of apoptosis, but rather an effect of apoptosis.

Capabilities that will enable them strengthen their leadership position include:

1.  Uses a cell permeant probe that can easily penetrate tissues and cells.
2. Very sensitive.
3. Specific, no reported false positives.
4. It is a direct measurement of an intracellular process of apoptosis, detects only active caspases and caspase active cells are always apoptotic.
5. Passage through the blood-brain barrier has been demonstrated.
6. Passage through the blood-retinal barrier has been demonstrated.
7. No background problems when injected intravenously.
8. Detects very early through late stage apoptosis.

ICT is continuing to invest heavily in developing new capabilties. Gary highlighlighted some of the breakthroughs that are on the horizon. I plan on announcing these as they become public.Stay tuned.