I am looking for an analitic column to evalute the hidrophobicity profile of different antibodies and its variants. High resolution and high selectivity it is need, since am I interested in really small diferences between hidrophobicity profile.
Protein-A affinity purification is the industry standard for mAbs, yielding up to 98% purity with one pass. You can check out this paper for details on the methodology: Article Heterologous recombinant expression of non-originator NISTmAb
You may consider columns from
Thermo http://www.thermofisher.com/order/catalog/product/088644
Phenomenex https://www.phenomenex.com/Application/Detail/19776?returnURL=/Application/Search&fsr=1
Agilent https://www.agilent.com/cs/library/applications/5991-0895EN.pdf
Merck https://www.sigmaaldrich.com/technical-documents/articles/analytical-applications/hplc/uhplc-analysis-of-a-monoclonal-antibody-system-suitability-standard-g1007502.html
For Analytic purposes HPLC SEC gives excellent resolution for antibodies and fragments, but if you expect differences in hydrophobicity only you should consider analytical HIC columns instead. With the amount of salt added, you can adjust resolution as required. In general, you should prefer small beads over large ones and if available, HPLC over FPLC as mentioned. Good luck !
I should add that my suggestion was assuming that your mAbs are from ascites or cultured cell stocks. Protein A & G bind selectively and with high affinity to the Fc region of IgG-type monoclonal and polyclonal Abs and fragments.
If you need to purify Abs raised in animals then there are several types of columns (ex: Sepharose CNBr/SH) that readily couple peptides, proteins (or protein domains), and haptens to the stationary phase. Immobilizing your Ab target is the most simple and effective way of purifying antigen-specific Abs in their native form. A comprehensive experimental M&M for several types of Ab purification can be seen here: http://doi.org/10.1007/978-1-59745-324-0_5
Your project sounds very interesting but I'd be surprised if the resolution of hydrophobic interaction chromatography is good enough to distinguish between different antibodies. Typically, hydrophobic interactions are increased by high salt concentrations so antibodies are applied to columns (such as hydroxyapatite, etc.) in high salt and antibodies can be eluted with a decreasing salt gradient. You may want to consider an alternative approach however. The binding of hydrophobic dyes such as ANS might prove to be more sensitive for your project?
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