title: Quantum dot-antibody conjugates via carbodiimide-mediated coupling for cellular imaging.
abstract: This chapter describes the processes of antibody (Ab) production, purification, conjugation to quantum dots (QDs), and the use of the conjugates produced in intracellular imaging of cell components and structures. Specifically, information is provided on the conjugation of carboxyl surface-terminated QDs to Abs via a one-step reaction using the water-soluble carbodiimide, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC). The chapter details the process of conjugate optimization in terms of its final fluorescence and biological activity. The method described should guarantee the production of QD-Ab conjugates, which outperform classic organic fluorophore-Ab conjugates in terms of both image definition produced and the longevity of the imaging agent.
authors.affiliation: Royal Veterinary College, Royal College Street, London, NW1 0TU, UK.
authors.forename: Daniel Alistair
authors.initials: DA
authors.lastname: East
authors.forename: Michael
authors.initials: M
authors.lastname: Todd
authors.forename: Ian James
authors.initials: IJ
authors.lastname: Bruce
mesh_terms: D000588:Amines / Q000737:chemistry; D000906:Antibodies / Q000737:chemistry* / Q000302:isolation & purification; D002234:Carbodiimides / Q000737:chemistry*; D018797:Cell Cycle Proteins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D042541:Intracellular Space / Q000378:metabolism; D057054:Molecular Imaging / Q000379:methods*; D045663:Quantum Dots / Q000737:chemistry*; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D014462:Ultrafiltration; D014867:Water / Q000737:chemistry
chemical_list: D000588:Amines; D000906:Antibodies; D002234:Carbodiimides; C410737:Cdc8 protein, S pombe; D018797:Cell Cycle Proteins; D029702:Schizosaccharomyces pombe Proteins; D014867:Water