Protein Arrays for Multidrug-resistance in Human Leukemia Cell Determination

Feng Gao,Xiaoxing Xu,Chunxiu Zhang,Baoan Chen,Zuhong Lu,Juan Du,Jian Cheng

doi:10.3390/s5040259

Feng Gao, Xiaoxing Xu + Show 5 more

Open Access

https://doi.org/10.3390/s5040259

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Abstract

A novel technique was developed, that was high throughput simultaneousscreening of multiple resistance protein expression based on a protein array system. Themethod combined the advantage of the specificity of enzyme-linked immunosorbentassays with the sensitivity and high throughput of microspot. In this system, the multipleresistance protein arrays were created by spotting the captured antibodies onto the glassslide. The arrays were then incubated with cell samples of leukemia patients. The boundproteins were recognized by biotin-conjugated antibodies and detected by CCD.Experiments demonstrated that three multiple resistance proteins, including Pgp, MRPand BCRP which are members of the ATP-binding-cassette (ABC) superfamily ofmembrane transporters could be simultaneously detected using this new approach.Research work shows the result is coincident with flow cytometry (FCM) (P>0.01). Itprovided a methodology to develop many high-density protein array systems to detect avariety of proteins. The protein arrays will provide a powerful tool to identify theleukemia cell protein expression and rapidly validate their MDR determination.

Highlights

The emergence of multidrug resistance (MDR) plays a crucial role in the failure of chemotherapy of leukemia patients [1,2]
One of the most common mechanisms implicated in causing MDR is in the multidrug proteins P-gp, MRP1, and BCRP — all belong to the ATP-binding cassette (ABC) transporter family, which is the ATP dependent, transmembrane drug efflux pump [3]
Three monoclonal antibodies of P-gp, MRP1 and BCRP were immobilized on a modified glass slide

Summary

Introduction

The emergence of multidrug resistance (MDR) plays a crucial role in the failure of chemotherapy of leukemia patients [1,2]. One of the most common mechanisms implicated in causing MDR is in the multidrug proteins P-gp, MRP1, and BCRP — all belong to the ATP-binding cassette (ABC) transporter family, which is the ATP dependent, transmembrane drug efflux pump [3]. Accurate determination of these MDR proteins is necessary as they may have important clinical implications. There is a strong interest and need for sensitive and rapid determination methods for MDR of leukemia cells to provide in-time warnings, low sample volume, and low cost and facilitate early clinical reverse treatment. Leukemia cells were incubated with the protein array, and detected and imaged by a CCD

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Conclusion