How can I transform a liquid bacteria protein (DNA or other ) into a solid state?
is there a method suitable for preserving protein structure without changing it?
for use in immunization lab animals. and analysis of protein structure by XRD to contact with immune cells by imitation program.
Dear Thamer Jaddoa Shihab Ahmed
sorry i do not undestand your question: Do you mean how you can preserve for long time purified recombinant proteins? or also DNA?
Regarding proteins, for animal immunizzation the long terms storage approach are:
1) Frozen at -20°C (with or with-out addiction of glycerol as cryoprotectant, ,15% of glycerol is enough in general). In case of glycerol addiction, i recommend you to store the protein at higher concentration (at least 10X) respect the required for the immunization, so glycerol % could be reduced at the moment of animal somministration.
2) Loiofilization
Theoretically liofilizzation have to be perfromed in absence of salts so in water or in (water/organic solvent) mixtures since the salts are highly concentrated during liofilizzation and can induce protein unfolding, aggregation, precipitation. But since not all the proteins are stable in water and since some proteins (e.g antibody) are stable also in presence of high salt concentration. a speifici liofilizzation protocol need to be settet-u protein by protein .
I suggest to you in all cases (freezing or liofilization) to perform a preliminary test (in a small lot of sample) about the effect of dose treatments can have in the protein folding.
In general i like to use DSF for this purpose.
in the following link of my blog you can see a small video abour the use of DSF to assess the effect of freezing in protein folding:
https://www.blogger.com/video.g?token=AD6v5dzr9KqZslTx9NeEjXx1XyPti8dU15eqYoE9TT8OEDaZQcNINuDKwzCAzVXkFWZA0btipKTr81doABh-VjpprwCcRukVcLhAXE_T5BzPxYbeQQA7VMFmc4DdpjS4XbOfxkwAhIWu
Regariding DNA: DNA is in general stable for long terms frozen at -20°C in water, otherwise you can perform lyofilizatio since the DNa is stable in water.
good luck
Manuele
Yes, I would like to obtain proteins (receptors or toxins) by doubling the DNA strand with a PCR machine, and after that I want to convert the protein, which is in liquid form (is it possible to photograph the shape of the protein in this state) into a powder (to keep it from changing) so that the animals are immunized?
Manuele Martinelli
Yes, I would like to obtain proteins (receptors or toxins) by doubling the DNA strand with a PCR machine, and after that, I want to convert the protein, which is in liquid form (is it possible to photograph the shape of the protein in this state) into a powder (to keep it from changing) so that the animals are immunized?
Dear Thamer Jaddoa Shihab Ahmed
it seems to me that you miss some steps. DNA and proteins are not the same thinkgs, you need to do some work to pass from DNA to protein
To produce the protein to be immunized in the mice, you have to amplify the dna by PCR, clone it in a vector for protein expression in some host as e.coli and/or mammalian and at this point try to express and purify the protein from this organism. The purified protein that could be used for immunize animals is in general stored at 4°C or -20°C (for long terms) in a buffer as PBS or Tris and in some cases with the addiction of glycerol for storage at -20°C
Alternative, you can try to immunize animals with DNA or mRNA (the same tecnology applied for the covid vaccines). In this way you do not have to purifity the proteins since the protein is produced by the cells of the animals, but the preparation and formulation of matherial is something not so simple, you need to have specific experience.
best
Manuele
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