minicircle – Minicircle

Experimental Vivisection — N6 — Version2

An update on version 2 of the N6 plasmid experiment. This is a living document that will be updated as we get new data.

This experiment is intended as a proof of concept for a possible future therapy. This experiment is in no way intended to cure or treat HIV: a cocktail of similar therapies, each delivering different antibodies, would be needed for a durable treatment.

The production of antibodies using plasmids, if feasible, could greatly impact patient outcomes.

Context This ‘Vector A’ was designed to express the gene for N6, an antibody that binds to the GP120 protein on HIV’s envelope. The binding action of N6 neutralizes the virus’s ability to infect cells, acting as both a prophylactic and treatment for ...

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Electrophoresis Gel Bands

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The Story Behind MiniCircle

In 2019 Tristan Roberts self-tested an experimental gene transfer mechanism created by biohackers.

We are developing the first open-source horizontal gene transfer protocol for humans. We are starting with genes that confer immunity to disease — broadly neutralizing antibodies. Although we are still in proof of concept stages (with accurate data expected in February 2020) we believe this will be the first functional vaccine for HIV.

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Vaccine Concept

Minicircles are small loops of DNA designed to express proteins for longer and more intensely than traditional plasmids.

Antibodies are large, Y-shaped proteins generated by the immune system to neutralize pathogens. In November 2016 National Institutes of Health discovered N6 and N6-LS, the first broadly neutralizing antibody (bnAbs) against HIV, capable of neutralizing 98.5% of HIV strains. Since then, more effective bnAbs have been found, capable of neutralizing >99.9% of HIV strains.

We incorporate a cocktail of three minicircles expressing three bnAbs.

DNA is complexed with PEI before injection. When transformed in vivo, the vaccine should confer immunity to HIV.

Experiment

To begin we started with injecting a single minicircle coding for N6. Blood is drawn every week. Because N6 is only capable of neutralizing 98.5% of HIV particles, 1.5% will remain as "escape variants" and this injection cannot cure HIV.

To do the Western Blot and ELISA we need the N6 Antibody Standard, N6 Fab fragment, and N6 anti-idiotype antibody.

Antibody Standard
Fab fragment
anti-idio type antibody

To verify antibody production we do the Western Blot (cheaper).

To quantify antibody production over time we do ELISA.

N6 Anti-ID antibodies bind predictably to N6. The N6 standard is a pure form of the N6 antibody which is intended to be produced in vivo. The Fab is a fragment of the N6 standard used to create the anti-ID antibodies.