TITLE

Novel antibody derivatives for proteome and high-content analysis

AUTHOR(S)
Schmidthals, Katrin; Helma, Jonas; Zolghadr, Kourosh; Rothbauer, Ulrich; Leonhardt, Heinrich
PUB. DATE
August 2010
SOURCE
Analytical & Bioanalytical Chemistry;Aug2010, Vol. 397 Issue 8, p3203
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
The understanding of cellular processes and their pathophysiological alterations requires comprehensive data on the abundance, distribution, modification, and interaction of all cellular components. On the one hand, artificially introduced fluorescent fusion proteins provide information about their distribution and dynamics in living cells but not about endogenous factors. On the other hand, antibodies can detect endogenous proteins, posttranslational modifications, and other cellular components but mostly in fixed and permeabilized cells. Here we highlight a new technology based on the antigen-binding domain of heavy-chain antibodies (VHH) from Camelidae. These extremely stable VHH domains can be produced in bacteria, coupled to matrices, and used for affinity purification and proteome studies. Alternatively, these VHH domains can be fused with fluorescent proteins and expressed in living cells. These fluorescent antigen-binding proteins called “chromobodies” can be used to detect and trace proteins and other cellular components in vivo. Chromobodies can, in principle, detect any antigenic structure, including posttranslational modifications, and thereby dramatically expand the quality and quantity of information that can be gathered in high-content analysis. Depending on the epitope chosen, chromobodies can also be used to modulate protein function in living cells. [Figure not available: see fulltext.]
ACCESSION #
52532552

 

Related Articles

  • What's new in histochemistry and cell biology? Zuber, Christian; Ziak, Martin // Histochemistry & Cell Biology;Dec2001, Vol. 116 Issue 6, p553 

    The article focuses on latest developments in histochemical techniques and their manifold applications. Green fluorescent protein and its derivatives have become indispensable reagents in cell biology research. Microwaving of paraffin-embedded tissues for antigen retrieval represents its classic...

  • Confomational Analysis of Soluble Oligomers of GFP Tagged Prion Protein By Fluorescence Fluctuation Spectroscopy. Sakata, Hiroshi; Horiuchi, Motohiro; Takahashi, Izumi; Kinjo, Masataka // Current Pharmaceutical Biotechnology;Jan2010, Vol. 11 Issue 1, p87 

    The conversion of prion protein (PrP) from the monomeric cellular isoform to the oligomeric pathological isoform is a crucial event in the pathogenesis of prion diseases. To investigate oligomer formation of PrP, enhanced green fluorescent protein (EGFP)-tagged PrP (EGFP-PrP) without the...

  • Selective induction of astrocytic gliosis generates deficits in neuronal inhibition. Ortinski, Pavel I.; Jinghui Dong; Mungenast, Alison; Cuiyong Yue; Takano, Hajime; Watson, Deborah J.; Haydon, Philip G.; Coulter, Douglas A. // Nature Neuroscience;May2010, Vol. 13 Issue 5, p584 

    Reactive astrocytosis develops in many neurologic diseases, including epilepsy. Astrocytotic contributions to pathophysiology are poorly understood. Studies examining this are confounded by comorbidities accompanying reactive astrocytosis. We found that high-titer transduction of astrocytes with...

  • 900. Specific AAV Serotypes Stably Transduce Hippocampal and Cortical Cultures with High Efficiency and Low Toxicity. Royo, Nicolas C.; Vandenberghe, Luk H.; Johnston, Julie; Maronski, Margaret; Dichter, Marc A.; Wilson, James M.; Watson, Deborah J. // Molecular Therapy;Jun2006, Vol. 13, pS347 

    Cultures of dissociated neurons and astrocytes are frequently used by neurobiologists to study neuronal physiology and pathophysiology. However, current methods for gene delivery into neurons in vitro are limited by very low efficiency, short-term gene expression, toxicity and/or the requirement...

  • 900. Specific AAV Serotypes Stably Transduce Hippocampal and Cortical Cultures with High Efficiency and Low Toxicity. Royo, Nicolas C.; Vandenberghe, Luk H.; Johnston, Julie; Maronski, Margaret; Dichter, Marc A.; Wilson, James M.; Watson, Deborah J. // Molecular Therapy;Jun2006, Vol. 13, pS347 

    Cultures of dissociated neurons and astrocytes are frequently used by neurobiologists to study neuronal physiology and pathophysiology. However, current methods for gene delivery into neurons in vitro are limited by very low efficiency, short-term gene expression, toxicity and/or the requirement...

  • Development: Dogma overturned. Cesari, Francesca // Nature Reviews Molecular Cell Biology;Dec2008, Vol. 9 Issue 12, p924 

    The article provides information on a research about the factors in the development of the mouse embryo. It notes that an early embryo is composed of the epiblast (ectoderm) and visceral endoderm. It also discusses a research on the population of visceral endoderm cells, which were discovered as...

  • Nondestructive micro-patterning of proteinous occlusion bodies in water by femtosecond laser-induced mechanical force. Takahiro Kaji; Eiji Kotani // Biomedical Microdevices;Apr2007, Vol. 9 Issue 2, p105 

    Abstract  Some insect virus produces proteinous occlusion bodies named polyhedra, on which it is possible to immobilize functional proteins, such as green fluorescent proteins, antibodies, and growth factors, with keeping their biological activity. In this work, several kinds of polyhedra...

  • A33scFv Green fluorescent protein, a recombinant single-chain fusion protein for tumor targeting. Ulf Petrausch; Jens Dernedde; Vânia Coelho; Hossein Panjideh; Dietmar Frey; Hendrik Fuchs; Eckhard Thiel; P. Markus Deckert // PEDS: Protein Engineering, Design & Selection;Dec2007, Vol. 20 Issue 12, p583 

    Chemical conjugates of monoclonal antibodies with fluorophores or enzymes have long been used for diagnostic purposes and experimental therapeutic approaches. Recombinant technology allows for the design and expression of tailored genuine fusion proteins, providing defined molecules as to size,...

  • The Arabidopsis apyrase AtAPY1 is localized in the Golgi instead of the extracellular space. Schiller, Madlen; Massalski, Carolin; Kurth, Thomas; Steinebrunner, Iris // BMC Plant Biology;2012, Vol. 12 Issue 1, p123 

    Background: The two highly similar Arabidopsis apyrases AtAPY1 and AtAPY2 were previously shown to be involved in plant growth and development, evidently by regulating extracellular ATP signals. The subcellular localization of AtAPY1 was investigated to corroborate an extracellular function....

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics