Volume 9, Issue 4 (2018)
JMBS 2018, 9(4): 557-564 |
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Gheraat M, Sajedi R, Jalilian N, Shanesaz M, MirShahi M. One-step Conjugation Method for Antibodies with CdTe Quantum Dots Using Activated Dextran . JMBS 2018; 9 (4) :557-564
URL: http://biot.modares.ac.ir/article-22-24334-en.html
URL: http://biot.modares.ac.ir/article-22-24334-en.html
1- Biochemistry Department, Biological Sciences Faculty, Tarbiat Modares University, Tehran, Iran
2- Biochemistry Department, Biological Sciences Faculty, Tarbiat Modares University, Tehran, Iran, Tarbiat Modares University, Nasr Bridge, Jalal-Al-Ahmad Highway, Tehran, Iran , sajedi_r@modares.ac.ir
3- Research Laboratory, Chemical Analysis, Mobin Chemistry Azma Company, Tehran, Iran
2- Biochemistry Department, Biological Sciences Faculty, Tarbiat Modares University, Tehran, Iran, Tarbiat Modares University, Nasr Bridge, Jalal-Al-Ahmad Highway, Tehran, Iran , sajedi_r@modares.ac.ir
3- Research Laboratory, Chemical Analysis, Mobin Chemistry Azma Company, Tehran, Iran
Abstract: (4214 Views)
Aims: The use of semiconductor quantum dots (QD) nanoparticles with emission spectrum in the visible region as a marker in immunoassays provides the user with an opportunity to detect the desired agent without using advanced equipment. Accordingly, the aim of this study was to present a one-step conjugation method for antibodies with CdTe quantum dots, using activated dextran.
Materials and Methods: In this experimental study, CdTe nanoparticles were synthesized and the transmission electron microscope was used to study the morphology of the synthesized QD of CdTe and the size, concentration, and stability of the synthesized nanoparticles were evaluated. In order to stabilize the nanoparticles synthesized by BSA (Bovine Serum Albumin), they were coated and connected to antibodies with activated dextran. Immunosuppression tests were used to evaluate the conjugated antibodies.
Findings: Spot and spherical nature were completely evident in the morphology of nanoparticles. The difference in QD and dBSA-QD displacement from the agarose gel confirmed the formation of dBSA-QD and the same dilution spectrum from nanoparticles was obtained in the presence and absence of BSA. Connecting with dBSA, in addition to maintaining and improving the properties of the nanoparticle's diffusion led to the creation of diverse functional groups for the next steps of nanoparticle connection. The fluorescence emission of nanoparticles was higher in both coated with dBSA and conjugated with antibodies than free nanoparticles. By using antibodies connected to nanoparticles, the detection limit of 30ng for protein antigen was obtained as an eye.
Conclusion: In the conjugation process, in order to connect CdTe quantum dots to antibodies via dextran, by coating nanoparticles with a denatured BSA in addition to increasing the stability of nanoparticles, new functional groups are created on the surface of the nanoparticle.
Materials and Methods: In this experimental study, CdTe nanoparticles were synthesized and the transmission electron microscope was used to study the morphology of the synthesized QD of CdTe and the size, concentration, and stability of the synthesized nanoparticles were evaluated. In order to stabilize the nanoparticles synthesized by BSA (Bovine Serum Albumin), they were coated and connected to antibodies with activated dextran. Immunosuppression tests were used to evaluate the conjugated antibodies.
Findings: Spot and spherical nature were completely evident in the morphology of nanoparticles. The difference in QD and dBSA-QD displacement from the agarose gel confirmed the formation of dBSA-QD and the same dilution spectrum from nanoparticles was obtained in the presence and absence of BSA. Connecting with dBSA, in addition to maintaining and improving the properties of the nanoparticle's diffusion led to the creation of diverse functional groups for the next steps of nanoparticle connection. The fluorescence emission of nanoparticles was higher in both coated with dBSA and conjugated with antibodies than free nanoparticles. By using antibodies connected to nanoparticles, the detection limit of 30ng for protein antigen was obtained as an eye.
Conclusion: In the conjugation process, in order to connect CdTe quantum dots to antibodies via dextran, by coating nanoparticles with a denatured BSA in addition to increasing the stability of nanoparticles, new functional groups are created on the surface of the nanoparticle.
Subject:
Agricultural Biotechnology
Received: 2017/12/21 | Accepted: 2018/03/11 | Published: 2018/12/21
Received: 2017/12/21 | Accepted: 2018/03/11 | Published: 2018/12/21
References
1. Michalet X, Pinaud F, Bentolila L, Tsay J, Doose S, Li J, et al. Quantum dots for live cells, in vivo imaging, and diagnostics. science. 2005;307(5709):538-44. [Link] [DOI:10.1126/science.1104274]
2. Resch-Genger U, Grabolle M, Cavaliere-Jaricot S, Nitschke R, Nann T. Quantum dots versus organic dyes as fluorescent labels. Nat Methods. 2008;5(9):763-75. [Link] [DOI:10.1038/nmeth.1248]
3. Smith AM, Ruan G, Rhyner MN, Nie S. Engineering luminescent quantum dots for in vivo molecular and cellular imaging. Ann Biomed Eng. 2006;34(1):3-14. [Link] [DOI:10.1007/s10439-005-9000-9]
4. Sukhanova A, Venteo L, Devy J, Artemyev M, Oleinikov V, Pluot M, et al. Highly stable fluorescent nanocrystals as a novel class of labels for immunohistochemical analysis of paraffin-embedded tissue sections. Lab Invest. 2002;82(9):1259-61. [Link] [DOI:10.1097/01.LAB.0000027837.13582.E8]
5. Nie S, Xing Y, Kim GJ, Simons JW. Nanotechnology applications in cancer. Annu Rev Biomed Eng. 2007;9:257-88. [Link] [DOI:10.1146/annurev.bioeng.9.060906.152025]
6. Sukhanova A, Devy J, Venteo L, Kaplan H, Artemyev M, Oleinikov V, et al. Biocompatible fluorescent nanocrystals for immunolabeling of membrane proteins and cells. Anal Biochem. 2004;324(1):60-7. [Link] [DOI:10.1016/j.ab.2003.09.031]
7. Lei J, Ju H. Signal amplification using functional nanomaterials for biosensing. Chem Soc Rev. 2012;41(6):2122-34. [Link] [DOI:10.1039/c1cs15274b]
8. Liu C, Jia Q, Yang C, Qiao R, Jing L, Wang L, et al. Lateral flow immunochromatographic assay for sensitive pesticide detection by using Fe3O4 nanoparticle aggregates as color reagents. Anal Chem. 2001;83(17):6778-84. [Link] [DOI:10.1021/ac201462d]
9. Mahmoud W, Rousserie G, Reveil B, Tabary T, Millot J, Artemyev M, et al. Advanced procedures for labeling of antibodies with quantum dots. Anal Biochem. 2011;416(2):180-5. [Link] [DOI:10.1016/j.ab.2011.05.018]
10. He Y, Sai LM, Lu H, Hu M, Lai W, Fan Q, et al. Microwave-assisted synthesis of water-dispersed CdTe nanocrystals with high luminescent efficiency and narrow size distribution. Chem Mater. 2007;19(3):359-65. [Link] [DOI:10.1021/cm061863f]
11. Xia Z, Xing Y, So MK, Koh AL, Sinclair R, Rao J. Multiplex detection of protease activity with quantum dot nanosensors prepared by intein-mediated specific bioconjugation. Anal Chem. 2008;80(22):8649-55. [Link] [DOI:10.1021/ac801562f]
12. So MK, Loening AM, Gambhir SS, Rao J. Creating self-illuminating quantum dot conjugates. Nat Protoc. 2006;1(3):1160-4. [Link] [DOI:10.1038/nprot.2006.162]
13. Mazumder S, Dey R, Mitra MK, Mukherjee S, Das GC. Review: Biofunctionalized quantum dots in biology and medicine. J Nanomater. 2009;2009:815734. [Link] [DOI:10.1155/2009/815734]
14. Nabiev I, Sukhanova A. Fluorescent colloidal particles as a detection tools in biotechnology systems. In: Eliassari A, editor. Colloidal Nanoparticles in Biotechnology. New York: Wiley & Sons Inc; 2008. [Link] [DOI:10.1002/9780470258552.ch6]
15. Mamedova NN, Kotov NA. Albumin-CdTe nanoparticle bioconjugates: Preparation, structure, and interunit energy transfer with antenna effect. Nano Lett. 2001;1(6):281-6. [Link] [DOI:10.1021/nl015519n]
16. Peng C, Li Z, Zhu Y, Chen W, Yuan Y, Liu L, et al. Simultaneous and sensitive determination of multiplex chemical residues based on multicolor quantum dot probes. Biosens Bioelectron. 2009;24(12):3657-62. [Link] [DOI:10.1016/j.bios.2009.05.031]
17. Jamieson T, Bakhshi R, Petrova D, Pocock R, Imani M, Seifalian AM. Biological applications of quantum dots. Biomaterials. 2007;28(31):4717-32. [Link] [DOI:10.1016/j.biomaterials.2007.07.014]
18. Yu W, Qu L, Guo W, Peng X. Experimental determination of the extinction coefficient of CdTe, CdSe, and CdS Nanocrystals. Chem Mater. 2003;15(14):2854-60. [Link] [DOI:10.1021/cm034081k]
19. Wang Q, Kuo Y, Wang Y. Shin G, Ruengruglikit C, Huang Q. Luminescent properties of water-soluble denatured bovine serum albumin-coated CdTe quantum dots. J Phys Chem B. 2006;110(34):16860-6. [Link] [DOI:10.1021/jp062279x]
20. He F. Laemmli-SDS-PAGE [Internet]. Stanford: Bio-protocol; 2011 [cited 2015 May 20]. Available from: https://bio-protocol.org/bio101/e80. [Link]
21. Charbgoo F, Mirshahi M, Sarikhani S, Saifi Abolhassan M. Synthesis of a unique high‐performance poly‐horseradish peroxidase complex to enhance sensitivity of immunodetection systems. Biotechnol Appl Biochem. 2012;59(1):45-9. [Link] [DOI:10.1002/bab.58]
22. de Menezes F, de Azevedo W, Alves JrS. A simple and fast synthesis route for preparing CdTe quantum dots in aqueous medium 11th Internatioal Conference on Advanced Materials. Rio de Janeiro. [Link]
23. Hermanson GT. Bioconjugate techniques. New York: Academic Press; 2013. [Link]
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