Additionally are there any other tips you could give in order for the right steps do be taken for the yield to be 100 percent and the enrichment value to be 1.0. Thanks in advance!
Jose: There is no 100% yield whenever you pass your sample through any column, including gel filtration resin. One key factor to take into account is to keep the protein of interest as active/native as possible and from potential degradation by proteases in the buffer/column. So, you should do some preparations to keep your system sterile, have protease inhibitors and other protective agents in the buffer and run columns at low temp as possible. These are the basics. Good luck.
I would advise you to start with the richest possible source of the protein, Getting high purity and yield is much easier when you start with a protein sample that has a lot of your protein in it, as from an overexpression system. If you have the capability to make an expression construct, include an affinity tag, allowing you to use affinity chromatography as part of the purification. Finally, make sure you have a good way to follow the protein through the purification, so you know which fractions contain it and in what amount.
Bear in mind that yield and purity are usually in opposition to each other. Think ahead about how much of the protein you need and how pure it needs to be.
If you know sequence of your protein and organism you can take many information from the sequence analysis. When you know pI of the protein you can use cation or anion exchange, you can try precipitation with Ammonium sulphate or if it comes from thermostable organism you can use also this feature. Overexpression (not native protein source) I would suggest with tag (his-tag, strep-tag, CBP tag or others). Affinity tag can significantly improve protein enrichment after one step and also improve solubility. When you have some amount of your protein after first try of purification, and it's not stable enough you can always try DSF (differential scanning fluorimetry) which can tell you which conditions are the best for your protein.
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