Talk:Protein purification

Untitled
Someone looking this up will likely want links to related entries on biochemical techniques. I believe that this article should first describe the concept of enrichment and methods of evaluating purity. Then it should provide a molecular property-based classification of techniques. Much work has to be done! --Bongotastic 13:42, 14 November 2005 (UTC)


 * We might as well get started. The only question is, should we make it an official science collaboration. I think that would help bring in more people to the task. But maybe there should be a start made before lots of people drop in. --JWSchmidt 16:26, 14 November 2005 (UTC)


 * Why not, if we organize the entry in such as way that the contribution can be broken down into small modules, the task woulnd't be daunting and people will probably join in. The organization I proposed isn't bad, but after looking at my Voet & Voet textbook, it think it can be improved too. To start the collaboration, I think the entry needs activity more than votes! --Bongotastic 19:14, 14 November 2005 (UTC)

Started to fill in the blanks, it give something to work with. --Bongotastic 04:09, 15 November 2005 (UTC)

I am new to this site, but thought I would add in some text on affinity and ion exchange chromatography. Feel free to change at will. :) MarcieM 15:39, 15 December 2005 (UTC)

References!!
If we want to make this a good article, it will definetly need references. Currently there are none. Please see if there is a book on your shelf that you can use to support any particular part of this article. It's harder to go back later and find references for a 'finished' article. ike9898 15:14, 28 February 2006 (UTC)
 * Hooray... The article just got it's first references... I just threw in some somewhat random reviews in some of the sections. This subject is so basic, that you could reference 1000's of different sources for the same stuff.... Of someone had a good purification text book, we could probably reference it all to that.... Kjaergaard 05:38, 4 March 2006 (UTC)

Overall structure
I don't really like the overall structure of the article at the moment.. THe whole "Separation on the basis of..." structure gives some problems since some techniques separate on the basis of several things at the same time.. And it means that all the chromatographic techniques gets spread out all over the place. I would like to have a section called chromatographic techniques etc... Any thoughts anyone..? Kjaergaard 04:08, 1 March 2006 (UTC)
 * OK, I was bold and just did it..... Kjaergaard 07:07, 3 March 2006 (UTC)

Centrifuge pictures
On commons:

Samsara contrib talk 16:35, 1 March 2006 (UTC)

Concentration
There is a concentration technique where you put your soln in a dialysis bag and surround the bag with PEG or something like that; water comes out but the PEG can't go in. Don't know what this is called or any more details; but the article should mention this. ike9898 18:53, 3 March 2006 (UTC)

Suggestions for Additional Topics
1) Ultrafiltration/Diafiltration steps are an integral part of most purification processes.  I think we should include a section on this topic (other than just the final concentration section).  Buffer exchanges and/or desalting steps are often needed.

2) A discussion on methods for monitoring purity (in addition to process yield) and a list of various assays that are common.

3) Perhaps we could add an example of a complex purification scheme...something common (and well published) like insulin maybe?  Any thoughts?

Additionally...you may want to label the picture of the AKTA as such...give credit to our oh-so-loved vendors. And the caption is a bit misleading as it says it is used for SEC, when in fact it can be used for any bench top chrom work. Just my two cents. :)

--MarcieM 01:25, 27 May 2006 (UTC)


 * Regarding the picture of the AKTA: I disagree. The brand of the specific piece of equipment is not important in this context, and it would be unnecessary POV to mention one specific manufacturer of chromatographic equipment. I changed the caption a bit, so it is clearer, that it can be used for other forms of chromatography. 1) Sure, just be bold and add it... 2) We should probably expand on the use of SDS-PAGE. Kjaergaard 05:23, 27 May 2006 (UTC)

Shakeflasks
The picture of the shakeflask is shakeflask.jpg ; the string shakeflask doesn't appear in the text. Why do I care about that? Because shakeflasks don't work; supposedly they are for "aeration" but I can assure you that they don't oxygenate. Before you set your mind to answering that, or thinking about it, ask yourself, "Have I ever measured the oxygen concentration in a shake flask?" and your answer will be no. If you answer is yes, please get in touch with me directly. Richard8081 14:31, 16 June 2007 (UTC)


 * Baffled flasks do, however, increase the growth rate of some organisms - whether this is a 02 concentration-level issue or a nutrient-distribution issue is debatable. As an illustration of a technique used to generate high cell counts (and thus high levels of the protein of interest), I think the image is reasonable. -- MarcoTolo 19:25, 17 June 2007 (UTC)

It's an illustration of a technique that doesn't work as described in its own Materials and Methods section, usually; it's an image of a common systematic error in biological research. Nothing reasonable about that. Richard8081 05:56, 17 July 2007 (UTC)

It appears to me that the picture is a flask of minimal media, rather than the more common 2xTY or LB broth. It would be nice to have an image of the more common growth medium, or at least label the current image as minimal medium (if that is indeed the case). Does anyone have an alternative picture? Sarahburge (talk) 21:09, 19 February 2011 (UTC)

Wiki Education assignment: Bio 401 Cell Biology with lab F2022
— Assignment last updated by Yeti2021 (talk) 20:21, 12 October 2022 (UTC)

Suggestions to elaborate on:

Lead Section: include "protein of interest" and "contaminants" phrase/word which are highly used in Protein Purification Biochemistry

Purpose: Discuss "protein purification scheme" and give simple easy to understand examples which can be elaborated on in the "Evaluating purification yield" section.

Preliminary Steps: Extraction: update "centrifugation" with "differential centrifugation" and explain process that leads to more discriminating purification techniques that are discussed further in the article. Also, include a link for an image on this.

Move "Precipitation and differential solubilization" under "Purification Strategies"

Before going into these strategies/techniques, restate that proteins can be purified according to solubility, size, charge, and binding affinity (which was mentioned in the lead section) but will bring clarity to each strategy mentioned. Add more citations: as previously mentioned by others in the talk — Preceding unsigned comment added by Yeti2021 (talk • contribs) 01:24, 21 October 2022 (UTC)

This was an educational Wikipedia assignment from Bio 401 Cell Biology with lab F2022. The published changes entailed minor formatting changes, such as moving sections in "Preliminary Steps" and "Purification Strategies" for a better flow, and updating headings under "Purification strategies". In addition, more information and definitions were added for clarity ("Protein purification"/"Purpose"/"Extraction"/"Ultracentrifugation"/ "Precipitation and differential solubilization"/"Size exclusion"/"Separation based on charge"/"Affinity chromatography"), and an up-to-date citation was included under "Purpose". — Preceding unsigned comment added by Yeti2021 (talk • contribs) 01:27, 28 November 2022 (UTC)