c19

search for a cure

What I would like to do

I want culture, grow, and genetically modify recombinatory antibody HEK 293 human cells such that S309 antibodies are produced by using the Cas9 protein known as CRISPR. I want to document and share this experience.

Why?

Cas9 is known to be used to cut and paste DNA. HEK 239 human cells are widely available (from every individual) and will reproduce quickly. S309 has been identified to be a functional anitbody to neutralize the COVID-19 virus (see NIH paper). By documenting and sharing the process of self-culturing and modifying HEK239 cells into S309 antibodies, an "everyman's vaccine" can be produced in home by any consenting adult for their own personal one-month immunity to the COVID-19 pandemic. The anticipated results are herd immunity, re-established trust in communities, and a reboot of economies potentially world-wide. This is how to protect loved ones, healthcare workers, and communities. This is the way 'back to normal'. This is the way.

Key Resources

Procedure

• DNA from whitepaper (David Ishee)

Growing and Storing HEK239

Proteins

• CaS9
• HEK239
• S309 Antibody

Cost Analysis

When?

What I know from https://www.dictionary.com/e/what-are-antibodies-and-do-they-kill-viruses/

• c19 is a pathogen
• immunity is indicated by the presence of antibodies
• antibodies are protein molecules produced by the immune system produced by the immune system in response to antigens
  • how are antibodies produced
  • what produces antibodies
• antigens are pathogens
• antibodies latch on to antigens using a unique bonding site
  • what is the name of this site
  • do the antibodies fit the antigen shape?
• antibodies are made by "B cells" aka "B lymphocytes"
• B cells are made in the marrow and found in the blood and lymph
• Antibodies have a distinctive "Y" shape which is key to how they work
• at the tips of antibodies are unique sites where they bind with a matching site on antigens and destroy them
  • what makes this site unique?
  • how does the antibody bond with the antigen?
  • why and how are the antigens destroyed by the antibody?
  • what is the defining structure of an antibody that destroys a common flu antigen?
  • what is the defining structure of the common flu antigen?
• antibodies (Ab) are also called immunoglobulins (Ig) and are found in the plasma of blood and certain cell membranes
• there are five types of Ig defined by location and function (see link above)
• IgG and IgM are the key players in terms of warding off infectious diseases...

What needs to happen

Phase 1

• Identify a common known antibody/antigen pair such that the antibody destroys the antigen
• define the structure and characteristics of the antibody
• define the structure and characteristics of the antigen
• define the structure and characteristics of the cell in the bone marrow that produces the antibody
• define a target antigen to destroy
• simulate the antibody
• simulate the antigen
• simulate the modification and creation of the antibody
• observe the effect of the mutant antibody on the target antigen
• repeat simulation of the modification, creation, and effect of the antibody until the desired structure of the antibody is identified
• define the structure of the bone marrow cells required to create the mutated antibody
• define the steps needed to modify the antibody using CRISPR

Phase 2

• modify existing antibodies to form to the target antibody structure (using CRISPR)
• modify marrow using CRISPR

Phase 3

• test on a small controlled real world sample
• scale to production

Resources

Data

Simulator

NIH White Paper

CRISPR

Coronavirus Live Stats Information and Graphs