Evenin'!
From: Adam Gaertner
Date: March 11, 2020 at 6:16 AM EDT
To: Fauci, Anthony (NIH/NIAID)
Subject: Coronavirus bioweapon production method
Hello Anthony,
This is how the virus was created.
Intervirion Fusion. HIV-luc(ACE2)(500 ng of p24) was mixed with 1,000 ng of p24 of HIV-gfp particles incorporating ASLV-A envelope, SARS-CoV S protein, or both envelopes in PBS at 4°C for 30 min to allow binding. Virions were adjusted to the desired pH with 0.1 M citric acid. PBS, TPCK-trypsin (final concentration 10 ng/ml), CTSL, cathespin B (CTSB) (final concentrations 2 ng/ml) or CTSL buffer alone was then added. Recombinant CTSL (R&D Systems) was preactivated by incubation for 15 min at 10 ng/ml in 50 mM Mes, pH 6.0, on ice. Recombinant CTSB (R&D Systems) was preactivated in 25 mM Mes, 5mM DTT, pH 5.0, for 30 min at 25°C. After a 10-min incubation at 25°C, proteolysis was halted by the addition of 300 nl of DMEM10 containing leupeptin (25 ng/ml) and STI (75ng/ml). Virions were then incubated at 37°C for 30 min to allow membrane fusion. 100 nl of the virion mixture was added in quadruplicate to HeLa-Tva cells pretrated for 1 h with leupeptin (20 ng/ml). The cells were spin-infected and incubated at 37°C for 5 h.
Truth is heavy.