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United States Department of Agriculture

Agricultural Research Service

Title: Affinity Selection of Compounds in a Fluoroquinolone Chemical Library by on-Line Immunoaffinity Deletion Coupled to Column Hplc

Authors
item Holtzapple, Carol
item Stanker, Larry

Submitted to: Analytical Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: August 23, 1998
Publication Date: N/A

Interpretive Summary: Novel chemical methods are used to develop new drugs that can cure or prevent disease in food animals. Although these methods can generate large numbers of potential drugs, it is often difficult to pick out of the mass of compounds the ones that will actually prove to be useful. Therefore, a simple, rapid, automated method was developed that is capable of selecting only those chemicals that have desirable characteristics. Other compounds that are present in solution are removed during a washing step, while promising drug leads are captured for further analysis. The method has the potential to greatly diminish the amount of time needed to screen potential drug candidates, thereby speeding up the drug discovery process.

Technical Abstract: To demonstrate that members of a fluoroquinolone chemical library can be separated on the basis of relative affinities for an antibody, an automated column switching system was used incorporating on-line immunoaffinity deletion coupled with hydrophobic interaction chromatography (HIC). A monoclonal antibody against sarafloxacin (Mab Sara-95) was covalently cross-linked to a protein G column and used to capture injected fluoroquinolones. A pH gradient in the mobile phase eluted the fluoroquinolones from the IAC column to the HIC column according to their relative affinities for the antibody, and final separation of the fluoroquinolones was accomplished on the HIC column before fluorescence detection. Since the fluoroquinolones clearly eluted from the IAC column on the basis of their relative affinities for the antibody, compounds exhibiting higher or lower relative affinity for the antibody could be selectively deleted from the library by exploiting the column switching capabilities of the system. The method described here is the first to report ¿immunoaffinity deletion¿ as a means to selectively extract chemicals with the desired binding characteristics from a mixture of closely related compounds.

Last Modified: 7/30/2014
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