Detection of Intracellular Proteins and Biomarkers Using Modified Silica Nanoparticles and Flow Cytometry In vitro

Abstract

In this paper, antibody-modified silica nanoparticles were successfully synthesized, and their average di-ameter was (109±9) nm. These particles were mixed with cell extracts to target a protein, then, an antibody labeled with fluorescein isothiocyanate(FITC) was added to form FITC-labeled nanoparticle complexes and the product was analyzed using flow cytometry. The results confirm that the intracellular proteins and biomarkers were precisely and sensitivity detected by the novel method in vitro. The FITC intensity of Akt antibody-conjugated particles was in-creased ≥10-fold compared with that of control samples in MCF-7 cells. Furthermore, it can also simultaneously measure several proteins when modified with different antibodies.

Keywords

Silica nanoparticle / Flow cytometry / Intracellular protein / Biomarker

References

[1] Nalejska E., Maczynska E., Lewandowska M. A. Mol. Diagn. Ther., 2014, 18(3): 273.

[2] Lourenco A. P., Benson K. L., Henderson M. C., Silver M., Letsios E., Tran Q., Gordon K. J., Borman S., Corn C., Mulpuri R., Smith W., Alpers J., Costantini C., Rohatgi N., Yang R., Haythem A., Biren S., Morris M., Kass F., Reese D. E. Clin. Breast Cancer, 2017, 17(7): 516.

[3] Chatterjee M., Hurley L. C., Tainsky M. A. Gynecol. Oncol. Rep., 2017, 21: 37.

[4] Yu X., Yuan Y., Zhi X., Teng B., Chen X., Huang Q., Chen Y., Guan Z., Zhang Y. Exp. Ther. Med., 2015, 10(3): 1175.

[5] Smits M., Mehra N., Sedelaar M., Gerritsen W., Schalken J. A. Expert Rev. Mol. Diagn., 2017, 17(8): 791.

[6] Moravec R., Divi R., Verma M. World J. Gastrointest Oncol., 2017, 9(6): 235.

[7] Salvadori M., Tsalouchos A. World J. Transplant, 2017, 7(3): 161.

[8] Hutchinson J., Jin J., Cardiff R. D., Woodgett J. R., Muller W. J. M. ol. Cell Biol., 2001, 21: 2203.

[9] Jin H., Wang X., Xin T., Gao P., Lin J. M., Liang S. Chin. Sci. Bull., 2008, 53(19): 2958.

[10] Gero E. J., Melsheimer R., Colcher D., Ferroni P., Giani S., Schlom J., Kaplan P. J. Clin. Lab. Anal., 1989, 3(6): 360.

[11] Sheng S. L., Wang Q., Huang G. Clin. Chim. Acta, 2007, 380(1/2): 106.

[12] Chen Y., Wang K., Liu Z., Sun R., Cui D., He J. Anal. Bioanal. Chem., 2016, 408(9): 2319.

[13] Fan H., Guo Z., Gao L., Zhang Y., Fan D., Ji G., Du B., Wei Q. Biosens Bioelectron, 2015, 64: 51.

[14] Zhang H. P., Han B., Jia Z. Z., Ding R. B., Xu B., Xu W. Q., Fan Z. M. Chem. J. Chinese Universities, 2017, 38(5): 860.

[15] Luo D., Han E., Belcheva N., Saltzman W. M. J. Control Release, 2004, 95: 333.

[16] Lu J., Liong M., Zink J. I., Tamanoi F. Small, 2007, 3: 1341.

[17] Kang M. S., Singh R. K., Kim T. H., Kim J. H., Patel K. D., Kim H. W. Acta Biomater., 2017, 55: 466.

[18] Gao X. H., Cui Y. Y., Levenson R. M., Chung L. W. K., Nie S. M. Nature Biotech., 2004, 22: 969.

[19] Liu Z., Davis C., Cai W., He L., Chen X., Dai H. Proc. Natl. Acad. Sci. USA, 2008, 105: 1410.

[20] Kim D., Park S., Lee J. H., Jeong Y. Y., Jon S. J. Am. Chem. Soc., 2007, 129: 7661.

[21] Ballou B., Lagerholm B. C., Ernst L. A., Bruchez M. P., Waggoner A. S. Bioconjugate Chem., 2004, 15: 79.

[22] Derfus A. M., Chan W. C. W., Bhatia S. N. Nano Lett., 2004, 4: 11.

[23] Poland C. A., Duffin R., Kinloch I., Maynard A., Wallace W. A., Seaton A., Stone V., Brown S., Macnee W., Donaldson K. Nature Nanotech., 2008, 3: 423.

[24] Zeng C., Shang W., Liang X., Liang X., Chen Q., Chi C., Du Y., Fang C., Tian J. ACS Appl. Mater. Interfaces, 2016, 8(43): 29232.

[25] An M., Li M., Xi J., Liu H. ACS Appl. Mater. Interfaces, 2017, 9(28): 23466.

[26] Vivero-Escoto J. L., Slowing I. I., Trewyn B. G., Lin V. S. Small, 2010, 6: 1952.

[27] Herzenberg L. A., Parks D., Sahaf B., Perez O., Roederer M., Herzenberg L. A. Clin. Chem., 2002, 48: 1819.

[28] Bonetta L. Nat. Meth., 2005, 2: 785.

[29] Wu Y., Campos S. K., Lopez G. P., Ozbun M. A., Sklar L. A., Buranda T. Anal. Biochem., 2007, 364: 180.

[30] Kapoor V., Hakim F. T., Rehman N., Gress R. E., Telford W. G. J. Immunol. Methods, 2009, 344: 6.

[31] Gu J., Fan W., Shimojima A., Okubo T. Small, 2007, 3(10): 1740.

[32] Lukacs S., Szabo N., Woodhams S. Case Rep. Urol., 2012, 2012: 724940.

[33] Tsou J. A., Kazarian M., Patel A., Galler J. S., Laird-Offringa I. A., Carpenter C. L., London S. J. Cancer Detect Prev., 2009, 32(4): 292.

[34] Ren H. T., Zhao Y. H., Guan H. Z., Li X. G., Shao Y. Q., Chen J. H., Chen L. Chin. J. Neuroimmunol and Neurol., 2011, 18(5): 332.

[35] Ma Z. X., Guan Y. P., Zhang X. X., Wang J., Wang B. C., Rong X. D., Hu H. Chin. J. Cancer Biother., 2015, 22(4): 500.