Hi
I would like to know the maximum ultra-centrifugation speed I could use that would still result in virus that is intact and able to invade cells. Is 35000g for 1hr at 4 degrees with STE buffer too aggressive?
Thank you
Dear Sir,
See these links may be useful for you.
1) Association Equilibrium of the HIV-1 Capsid Protein in a Crowded ...
www.ncbi.nlm.nih.gov › ... › PubMed Central (PMC) Traduire cette page
de R Bocanegra - 2013 - Cité 3 fois - Autres articles Polymerization of the intact capsid protein (CA) of HIV-1 into mature capsidlike ..... These s values were corrected to standard conditions (water, 20°C, and infinite ... After the equilibrium scans, a high-speed centrifugation run (48,000 rpm) was ...
2) A highly efficient and consistent method for harvesting large volumes ...
www.nature.com/gt/journal/v8/n22/.../3301587a.html Traduire cette page A transient, three-component, HIV-based vector system consisting of a ... Furthermore, because rotors that are suitable for low-speed centrifugation have a ... the PLL treatment had a 25% higher p24 value than the ultracentrifugation group, ...
I used ultra high speed to cushion HTLV and HIV, usually without a problem, and the speed depends a lot on the rotor you use. When I do larger volumes (8X50mL), I run 90% of the max rotor speed, which in my case is 33,300 for 2 1/2 hours. When I run smaller volumes (6X30mL) in a swinging bucket, I go with 27,000 for 3 Hours. Have not experienced infectivity issues using these parameters. We also pelleted lentivirus in a swinging bucket under the same conditions as HIV and again no problems. Hope that helps.
We routinely use ultra-centrifugation at 30,000 rpm without any cushion to concentrate HIV, without any loss in infectivity.
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