Researching GE's AKTA Pure HPLC unit for the detection of trace proteins in bovine milk. I am looking at getting a result every 5-10 minutes over a duration of 16 hours per day.
With that in mind, is there a better instrument for this purpose?
Thanks in advance for any advice.
Since milk already contains a lot of protein, including casein and immunoglobulin, I think it would be difficult to detect trace proteins by chromatography unless you could first selectively remove the major proteins. For high throughput, ELISAs might be a good choice if you have antibodies against the proteins of interest, and a standard for each.
Hello Mr. Rajendran!
Please try to find some ELISA method for minor proteins in milk ..if you are thinking of assessment of Ig or lactoferin I think that this is nice instrument ...here in Macedonia we have problems with lactoferin examinations and also HPLC procedure is very expensive, so you should check on internet or to ask in some laboratories about this detection.....good luck in your work!
Vesna
Thanks Adam and Vesna.
I am not entirely familiar with ELISA and a quick check has given me some background information. Not many systems are capable of quick analysis but AlphaLISA by Perkin Elmer might do the trick.
I am still open to different ideas such as ICP-MS/OES,
Hello,
I agree with Adam and Vesna that ELISA could be a good solution.The problem with milk proteins is where to get the GOOD ANTIBODIES and the good standards, as Adam said!
To my knowledge the AKTA systems are more for sample preparation and purification than for analytical purposes. I suppose this system could work quite well if you used it to eliminate the major proteins (caseins, alpha-LAC, beta-lG, Serum Albumin) in milk or whey on some Ion Exchange column, and then you can concentrate your sample and use it on some RP-HPLC system...
Otherwise I can suggest to just precipitate your caseins at pH 4.6 (with acetic acid-Na acetate) and then to get rid of beta-LG by salting out at pH 2. this would leave you with a concentrated fraction of the 10% minor proteins in milk (but with huge amounts of alpha-LAC and Serum albumin).
If money is not an issue for you, you can go for an LC-MS analysis of your minor proteins on monolyth columns, the run can be 5 to 10 minutes short and your identifications will be much better!
Good luck.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Proteins