Abstract
This study presents the development of targeted and antibody cross-linked QDs and explores whether these bioconjugates could specifically and effectively label Cu/Zn superoxide dismutase (SOD1) on fixed cells and tissues. QD-antibody conjugation was achieved by using our previously invented AmiNoacid (monomer) Decorated and Light Underpining Conjugation Approach (ANADOLUCA) method. In this method, we have used a photosensitive aminoacid monomer having ruthenium complex which is a synthetic and inexpensive material for the preparation of bioconjugates. Its specificity was demonstrated by extracting the active enzyme from rat liver lysate by using the bioconjugate. It provided accurate antibody orientation, high specificity and mechanic stability. The protocol steps for QD-antibody conjugation and specimen preparation were described in detail. The nanobioconjugates were prepared under mild conditions (for example in day light), independent of pH and temperature, without affecting conformation and function of protein. This protocol is simple, inexpensive and can be successfully adapted to detect other targets on different cell types and tissues.
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References
Bancroft JD, Gamble M (2002) Theory and practice of histological techniques. E.P. Churchill Livingstone, Edinburgh
Catimel B, Rothacker J, Nice E (2001) The use of biosensors for microaffinity purification: an integrated approach to proteomics. J Biochem Biophys Methods 49:289–312
Chang LY, Slot JW, Geuze HJ, Crapo JD (1988) Molecular immunocytochemistry of the Cu, Zn superoxide dismutase in rat hepatocytes. J Cell Biol 107(6):2169–2179
Dahan M (2006) From analog to digital: exploring cell dynamics with single quantum dots. Histochem Cell Biol 125:451–456
Deerinck TJ (2008) The application of fluorescent quantum dots to confocal, multiphoton, and electron microscopic imaging. Toxicol Pathol 36:112–116
Diltemiz SE, Say R, Büyüktiryaki S, Hür D, Denizli A, Ersöz A (2008) Quantum dot nanocrystals having guanosine imprinted nanoshell for DNA recognition. Talanta 75:890–896
Frangioni JV (2003) In vivo near-infrared fluorescence imaging. Curr Opin Chem Biol 7:626–634
Ghasemi Y, Peymani P, Afifi S (2009) Quantum dot: magic nanoparticle for imaging, detection and targeting. Acta Biomed 80:156–165
Giepmans BN (2008) Bridging fluorescence microscopy and electron microscopy. Histochem Cell Biol 130(2):211–217
Hür D, Ekti SF, Say R (2007) N-Acylbenzotriazole mediated synthesis of some methacrylamido amino acids. Lett Org Chem 4(8):585–587
Jamiesona T, Bakhshia R, Petrovaa D, Pococka R, Imanib M, Seifaliana AM (2007) Biological applications of quantum dots. Biomaterials 28:4717–4732
Kawamata H, Manfredi G (2008) Different regulation of wild-type and mutant Cu, Zn superoxide dismutase localization in mammalian mitochondria. Hum Mol Genet 17(21):3303–3317
Kulkarni SK, Ethiraj AS, Kharrazi S, Deobagkar DN, Deobagkar DD (2005) Synthesis and spectral properties of DNA capped CdS nanoparticles in aqueous and non-aqueous media. Biosens Bioelectron 21:95–102
Kumar SV, Jain R, Mokhiber K, Venezia A, Sheehan A, Spivack DS (2005) Exfoliated buccal and microdissected lung cell expression of antioxidant enzymes. Cancer Detect Prev 29:552–561
Ladha S, Mackie AR, Clark DC (1994) Cheek cell membrane fluidity measured by fluorescence recovery after photobleaching and steady-state fluorescence anisotropy. J Membr Biol 142:223–228
Matsumoto AO, Fridovich I (2001) Subcellular distribution of superoxide dismutases in rat liver: Cu, Zn-SOD in mitochondria. J Biol Chem 276(42):38388–38393
Mattoussi H, Medintz IL, Clapp AR et al (2004) Luminescent quantum dot-b, oconjugates in immunoassays, FRET, biosensing and imaging applications. J Assoc Lab Autom 9:28–32
Michalet X, Pinaud FF, Bentolila LL et al (2005) Quantum dots for live cells, in vivo imaging, and diagnostics. Science 307:538–544
Parak WJ, Pellegrino T, Plank C (2005) Labelling of cells with quantum dots. Nanotechnology 16:R9–R25
Say R (2009) Photosensitive aminoacid-monomer linkage and bioconjugation applications in life sciences and biotechnology. PCT/IB2009/055707 (applied for)
Skepper JN, Powell JM (2008) Immunogold staining of London resin white sections for transmission electron microscopy (TEM). CSH Protoc. doi:10.1101/pdb.prot5016
Smith AM, Duan H, Mohs AM, Nie S (2008) Bioconjugated quantum dots for in vivo molecular and cellular imaging. Adv Drug Del Rev 60:1226–1240
Stringer RC, Schommer S, Hoehn D, Grant SA (2008) Development of an optical biosensor using gold nanoparticles and quantum dots for the detection of porcine reproductive and respiratory syndrome virus. Sens Act B 134:427–431
Sweeney E, Ward TH, Gray N, Womack C, Jayson G, Hughes A, Dive C, Byers R (2008) Quantitative multiplexed quantum dot immunohistochemistry. Biochem Biophys Res Commun 374:181–186
Turkova J (1999) Oriented immobilization of biologically active proteins as a tool for revealing protein interactions and function. J Chromatogr B 722:11–31
Wang HZ, Wang HY, Liang RQ, Ruan KC (2004) Detection of tumor marker CA125 in ovarian carcinoma using quantum dots. Acta Biochim Biophys Sin 36(10):681–686
Winterbourn C, Hawkins R, Brian M, Carrell R (1975) The estimation of red cell superoxide dismutase activity. J Lab Clin Med 85:337–341
Xing Y, Chaudry Q, Shen C et al (2007) Bioconjugated quantum dots for multiplexed and quantitative immunohistochemistry. Nat Protoc 2(5):1152–1165
Xu C, Xing B, Rao J (2006) A self-assembled quantum dot probe for detecting b-lactamase activity. Biochem Biophys Res Commun 344:931–935
Yoo J, Kambara T, Gonda K, Higuchi H (2008) Intracellular imaging of targeted proteins labeled with quantum dots. Exp Cell Res 314:3563–3569
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Say, R., Kiliç, G.A., Özcan, A.A. et al. Investigation of photosensitively bioconjugated targeted quantum dots for the labeling of Cu/Zn superoxide dismutase in fixed cells and tissue sections. Histochem Cell Biol 135, 523–530 (2011). https://doi.org/10.1007/s00418-011-0801-7
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DOI: https://doi.org/10.1007/s00418-011-0801-7