Protein arrays for multidrug-resistance in human leukemia cell determination

Abstract

A novel technique was developed, that was high throughput simultaneous screening of multiple resistance protein expression based on a protein array system. The method combined the advantage of the specificity of enzyme-linked immunosorbent assays with the sensitivity and high throughput of microspot. In this system, the multiple resistance protein arrays were created by spotting the captured antibodies onto the glass slide. The arrays were then incubated with cell samples of leukemia patients. The bound proteins were recognized by biotin-conjugated antibodies and detected by CCD. Experiments demonstrated that three multiple resistance proteins, including Pgp, MRP and BCRP which are members of the ATP-binding-cassette (ABC) superfamily of membrane transporters could be simultaneously detected using this new approach. Research work shows the result is coincident with flow cytometry (FCM) (P>0.01). It provided a methodology to develop many high-density protein array systems to detect a variety of proteins. The protein arrays will provide a powerful tool to identify the leukemia cell protein expression and rapidly validate their MDR determination. © 2005 by MDPI.