Re: [DIYbio] Re: help me setup DIY-BIO home Lab to produce Recombinant Factor VIII

On Tue, Feb 10, 2015 at 12:01 PM, Sunil Phani
<consultsunilphani@gmail.com> wrote:
> i may be sounding obnoxious but why can we hack a typical UV & RO (ultra
> violet and reverse osmosis ) water purifier off the shelf
> to achieve protein purification ... coz every high end water purifiers now a
> days promises to be having chromatography and filtration
> i wonder if there is already a DIYBIO chromatography using off the shelf
> UV & RO water purifier coz
> every other UV & RO water purifier discusses deeply how it separates organic
> and inorganic
> solvents and compounds from water.. then just as appetite to brain why cant
> we give it a
> thought on its merits
>
> pros
>
> every water purifier have chromatography and filtration mechanism

Generally the goal of water filters is to retain as many molecules of
non-water as possible, and to let water pass.


>
> every water purifier have pH, UV sensors, but i don't know weather they have
> conductivity sensors are

If it is a UV water filter, the UV sensor probably is looking at very
high signal-strengths, and even if the sensor is suitable, the
amplifier/analog-to-digital circuit is probably not sensitive enough
or in the right amplification range. UV sterilization lights will be
VERY bright relative to the amount of light absorbed by presence of an
analyte in solution.

>
> every water purifier have filtration mechanism ( may not be 2 µm)
>
> most of the UV & RO water purifier have column chromatography (suitable
> resin is arguable)

yes the resin in filters would probably be targeting non-polar
molecules, as water is polar... otherwise the resin may simply be
adding a size-exclusion filtration.

> most of the UV & RO water purifier have ion exchange chromatography (but we
> need to hack it, like we were hacking Hp inkjet printers to print 3d
> microbial cells )

It really depends on the type of filter... I bet there are some
filters that allow ions through no-problem, but are targeting things
like viruses, bacteria, pesticides, environmental toxins.


> needs some reverse engineering to achieve clarified lysate purification
>
> needs some reverse engineering to achieve HIC by adding ammonium sulphate
> etc, in recursively cycles
>
> most of the UV & RO water purifier have TFF but we need to hack permeate
> and retentate chambers and focus in recycling renantate several time through
> diafiltration for efficiency
>
>
> guys this needs some kind of hacking to off the shelf UV & RO water purifier
> pls give it a thought and come up with your ideas and viewpoints on this
> topic.,
> becausehow often can every DIYBIO labs can afford such whopping expensive
> proprietary chromatographys lets rip open an opensource platform for
> purifying
> proteins using off the shelf purifiers


Paper is a commonly available chromatography media, agarose and
gelatin are also available.

From a quick google, I get: "Some of the first ion exchangers used
were inorganic and made from aluminosilicates (zeolites). Although
aluminosilicates are not widely used as ion exchange resins used. "
from:
http://chemwiki.ucdavis.edu/Analytical_Chemistry/Instrumental_Analysis/Chromatography/Chromatographic_Columns

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