Typical protocols for lambda phage in vitro assembly call for you to dump all your viral components and buffer solutions into water. Water is a great solvent, drives extraction of energy from atp, and probably contributes structurally at various steps in the process. But I am curious to know: how anhydrous can you get and still have at least half the typical yield of fully formed phages? Is there an alternative solvent that can suspend and disperse the viral particles and buffer solution, and then water can be subsequently added as a solute to drive the reaction? Of course, I am googling away at this question. But maybe a person reading this knows or at least has heard of something like this? Thank you in advance!
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$\begingroup$ By anhydrous you mean mixing the same concentration of viral proteins in a polar solvent that is not water? What would you suggest? Methanol? $\endgroup$– Buck Thorn ♦Commented Oct 29, 2022 at 11:51
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$\begingroup$ @BuckThorn yes, some type of solvent that is not water, so that water can be subsequently added in varying degrees to drive assembly. Is there a paper written on this already? What other solvents are possible candidates besides methanol? $\endgroup$– edmund sheltoCommented Oct 29, 2022 at 12:08
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1$\begingroup$ Proteins can sometimes be dissolved in solvents other than water, but there is a risk of denaturation that can then lead to aggregation. Proteins aren't like lego pieces that remain in one uniform shape regardless of solvent. $\endgroup$– Buck Thorn ♦Commented Jan 20, 2023 at 7:55
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$\begingroup$ Good solvents for proteins are trifluoroethanol, phenol, and maybe HFIP? I think most of the time solubilization will also involve denaturation though. $\endgroup$– Curt F.Commented Jan 20, 2023 at 19:06
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$\begingroup$ Also for lambda phage assembly you need to solvate the genomic DNA, too, right? It's much harder to imagine DNA as very soluble in anything that isn't water... $\endgroup$– Curt F.Commented Jan 20, 2023 at 19:07
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After a lot of research, I have narrowed it down to a few candidates: cyclohexane, methyl formate, 1-dodecanethiol, acetate, 1-Ethyl-3-methylimidazolium chloride, ethylammonium nitrate (EAN), ethanolammonium nitrate (EtAN), glycerol, and good old methanol. The last four are the most promising. Please note however that none of this has been confirmed yet through experiments.
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$\begingroup$ How on earth did you narrow it down without doing experiments? $\endgroup$– Curt F.Commented Jan 20, 2023 at 19:03
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$\begingroup$ Ummm.... exactly. I am in the design phase, narrowing down possible candidates before I go spending money. Any ideas? Thanks! $\endgroup$ Commented Jan 25, 2023 at 15:04