Intrabody

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In biochemistry and immunology, an antibody that binds intracellular proteins within a cell is referred to as intrabody (from intracellular and antibody ) .

properties

Since there are currently no effective mechanisms for transporting antibodies into a cell, it is necessary that the antibody in question is expressed as a recombinant protein in the target cell . This can be achieved in transgenic animals or through gene therapy . Therefore, antibodies that have been modified for intracellular activity are referred to as intrabodies , even if they were produced in bacteria or other cells that are not target organisms. The term can therefore be used for various forms of protein targeting. After expression, the antibody can remain in the cytoplasm , have a nuclear localization signal , or be introduced into the interior of the endoplasmic reticulum by means of a KDEL sequence by translocation during translation . Since antibodies are normally secreted, the signal sequences must be adapted through a protein design. For this purpose, single chain antibodies are used, the VL domains of the antibodies are changed to improve their stability, antibodies are selected that are more resistant to intracellular degradation mechanisms, or fusion proteins with protein tags ( e.g. with the maltose binding protein ), which are intracellularly more stable, are produced are. Such optimizations have produced stable intrabodies that have various properties and may be used against hepatitis B , avian influenza , prion diseases , inflammation , Parkinson's disease and Huntington 's disease . A knockdown of membrane proteins or secreted proteins can be achieved with ER intrabodies. While only selected antibodies are often functional in the cytosol, antibodies that are used as ER intrabodies can fold in their natural compartment, so that the optimizations required for cytosolic antibodies are not necessary.

Individual evidence

  1. ^ SY Chen, J Bagley, WA Marasco: Intracellular antibodies as a new class of therapeutic molecules for gene therapy . In: Human gene therapy . 5, No. 5, 1994, pp. 595-601. doi : 10.1089 / hum . 1994.5.5-595 . PMID 7914435 .
  2. ↑ AL Marschall, FN Single, K Schlarmann, A Bosio, N Strebe, J van den Heuvel, A Frenzel, S Dübel: Functional knock down of VCAM1 in mice mediated by endoplasmatic reticulum retained intrabodies . In: MAbs . 6, No. 6, 2014, pp. 1394-401. doi : 10.4161 / mabs.34377 . PMID 25484057 .
  3. ^ PA Cohen, JC Mani, DP Lane: Characterization of a new intrabody directed against the N-terminal region of human p53 . In: Oncogene . 17, No. 19, 1998, pp. 2445-56. doi : 10.1038 / sj.onc.1202190 . PMID 9824155 .
  4. ↑ AM Mhashilkar, J Bagley, SY Chen, AM Szilvay, DG Helland, WA Marasco: Inhibition of HIV-1 Tat-mediated LTR transactivation and HIV-1 infection by anti-Tat single chain intrabodies . In: The EMBO Journal . 14, No. 7, 1995, pp. 1542-51. PMID 7537216 . PMC 398241 (free full text).
  5. ↑ Yurong Yang Wheeler, Timothy E. Kute, Mark C. Willingham, Si-Yi Chen, and David C. Sane: Intrabody-based strategies for inhibition of vascular endothelial growth factor receptor-2: effects on apoptosis, cell growth, and angiogenesis . In: The FASEB Journal . 2003. doi : 10.1096 / fj.02-0942fje .
  6. ^ PA Cohen, JC Mani, DP Lane: Characterization of a new intrabody directed against the N-terminal region of human p53. . In: Oncogene . 17, No. 19, 1998, pp. 2445-56. doi : 10.1038 / sj.onc.1202190 . PMID 9824155 .
  7. ↑ A Auf Der Maur, D Escher, A Barberis: Antigen-independent selection of stable intracellular single-chain antibodies. . In: FEBS Letters . 508, No. 3, 2001, pp. 407-12. doi : 10.1016 / S0014-5793 (01) 03101-5 . PMID 11728462 .
  8. ↑ S Shaki-Loewenstein, R Zfania, S Hyland, WS Wels, I Benhar: A universal strategy for stable intracellular antibodies. . In: Journal of immunological methods . 303, No. 1-2, 2005, pp. 19-39. doi : 10.1016 / j.jim.2005.05.004 . PMID 16045924 .
  9. ↑ B Serruys, F Van Houtte, P Verbrugghe, G Leroux-Roels, P Vanlandschoot: Llama-derived single-domain intrabodies inhibit secretion of hepatitis B virions in mice . In: Hepatology (Baltimore, Md.) . 49, No. 1, 2009, pp. 39-49. doi : 10.1002 / hep.22609 . PMID 19085971 .
  10. ↑ MM Mukhtar, S. Li, W. Li, T. Wan, Y. Mu, W. Wei, L. Kang, ST Rasool, Y. Xiao, Y. Zhu, J. Wu: Single-chain intracellular antibodies inhibit influenza virus replication by disrupting interaction of proteins involved in viral replication and transcription. In: The international journal of biochemistry & cell biology. Volume 41, Number 3, March 2009, ISSN  1878-5875 , pp. 554-560, doi : 10.1016 / j.biocel.2008.07.001 . PMID 18687409 .
  11. ↑ I Filesi, A Cardinale, S Mattei, S Biocca: Selective re-routing of prion protein to proteasomes and alteration of its vesicular secretion prevent PrP (Sc) formation . In: Journal of Neurochemistry . 101, No. 6, 2007, pp. 1516-26. doi : 10.1111 / j.1471-4159.2006.04439.x . PMID 17542810 .
  12. ↑ N. Strebe, A. Guse, M. Schüngel, T. Schirrmann, M. Hafner, T. Jostock, M. Hust, W. Müller, S. Dübel: Functional knockdown of VCAM-1 at the posttranslational level with ER retained antibodies. In: Journal of immunological methods. Volume 341, Number 1-2, February 2009, ISSN  0022-1759 , pp. 30-40, doi : 10.1016 / j.jim.2008.10.012 . PMID 19038261 .
  13. ↑ C Zhou, S Przedborski: Intrabody and Parkinson's disease . In: Biochimica et Biophysica Acta . 1792, No. 7, 2009, pp. 634-42. doi : 10.1016 / j.bbadis.2008.09.001 . PMID 18834937 . PMC 2745095 (free full text).
  14. ↑ A Cardinale, S Biocca: The potential of intracellular antibodies for therapeutic targeting of protein-misfolding diseases . In: Trends in Molecular Medicine . 14, No. 9, 2008, pp. 373-80. doi : 10.1016 / j.molmed.2008.07.004 . PMID 18693139 .
  15. ↑ AL Marschall, S Dübel, T Böldicke: Specific in vivo knockdown of protein function by intrabodies . In: MAbs . 7, No. 6, 2015, pp. 1010-35. doi : 10.1080 / 19420862.2015.1076601 . PMID 26252565 .