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BIOASSAYS

Updated: Jun 4, 2019

I have developed a method to produce NAPE a precursor of NAE from natural sources. I have identified an anti-cancer compound present in a commercial product for a client that has some exciting prospects. I have developed a robust, rapid, real-time method for the detection of HDAC inhibition. I have developed a new model for Unified Field based on the principles of an electron that I use to determine minimized energy states in complex biological systems. I have identified the compound responsible for the anti-inflammatory activity in Manuka honey. I have identified an elastase I digest of epiphyseal cartilage that has apoptotic properties due to the presence of histone H2A derived peptide. I have identified a number of extracts that inhibit angiogenesis. I have extensive experience with chromatography and have developed new methods that allow the separation of proteins based on the conformation of a protein bound to the resin. I have identified numerous biological extracts that modulate inflammation for the Red meat Industry.

I have developed an ECM product from sheep stomach by developing the STOF process and the technique to remove the epithelium from the tissue. This is not based on osmosis. Endoform

I have also developed the peptide GAP573 that has wound healing and tissue regeneration potential through modulation of GAP junction modulation due to its homology to Cx-43 and the interaction with ZO PDX-2 domain. The peptide is taken from the C-terminal extension of beta B2 crystallin which is released by elastase I cleavage C-terminal to Val to release the peptide FHPSS. The cool thing about the process was the ability of elastase I to selectively cleave the peptide off the protein in the absence of calcium. So something unusual was observed. The peptide bound calcium and precipitated out of solution so I ended up using this potential to form a combination product that made the peptide release slowly from an alginate gel as the calcium was removed from the calcium peptide salt and into a calcium alginate gel over time. Funny how things work or at least how you think they work.


Love the biology of the human eye. The formation of a blood vessel feeding the developing lens in the embyro. The formation of the blood vessel is controlled by crystallin proteins within the eye lens. This causes the growth and regression of the blood vessel through angiogenesis. The peptide also has the properties of turning off inflammation due to its ability to stop inflammatory cells from talking to the endothelial cells so they cannot enter into tissue. Pretty cool seeing them trapped inside blood vessels. The epithelium remained undifferentiated for longer than usual as I did not know how long to keep applying the peptide alginate gel, however, this provided a good confirmation of the mode of action which was gap junction modulation. The cells grew faster and migrated more than usual providing the regenerative capacity to heal the tissue without limited scarring. Awesome! I have used it personally so know it helps looking forward to moving this one forward.



One the cool things I did during this work was to develop a bioassay using the actual tissue that I extracted the peptide from and this was an eye lens gap junction communication assay. Added fluorescein to a whole sheep eye lens and watched to see if the fluorescein migrated deep into the lens in the presence or absence of the peptide that I extracted. So the model worked and demonstrated that the peptide was able to modulate gap junction communication. Thanks for the insight Prof Colin Green from Auckland University. He was working on RNAi technology for the modulation of gap junction communication and he said that you needed technology to be better than what was currently available. Someone else also told me that anything would heal a wound. I said I agreed as long as it was not chronic. So how and why has been my life's work. I now understand and it's weird but cool.


#gap junction modulation #GAP573 #wound healing #tissue regeneration