logo SBA

ETD

Archivio digitale delle tesi discusse presso l’Università di Pisa

Tesi etd-09242012-000122


Tipo di tesi
Tesi di laurea magistrale
Autore
CHIRICHELLA, MICHELE
Indirizzo email
michele.chirichella@hotmail.it
URN
etd-09242012-000122
Titolo
In vivo selection of intrabodies for post-translational protein targeting
Dipartimento
BIOLOGIA
Corso di studi
BIOLOGIA MOLECOLARE E CELLULARE
Relatori
relatore Prof. Cattaneo, Antonino
controrelatore Prof. Andreazzoli, Massimiliano
controrelatore Prof.ssa Dente, Luciana
Parole chiave
  • antibody
  • bait
  • intrabodies
  • post-translational modification
  • protein silencing
  • protein targeting
  • PTM
  • scFv
  • single domain antibodies
  • yeast two-hybrid
Data inizio appello
18/10/2012
Consultabilità
Non consultabile
Data di rilascio
18/10/2052
Riassunto
Intracellular Antibodies (Intrabodies) are engineered antibodies (or antibody domains) ectopically expressed in several cell types to specifically perturb the function of intracellular antigens. (Biocca & Cattaneo, 1995)

Since Antibodies are proteins whose folding depends on the formation of critical disulphide bonds, the successful recognition of the physiological target, located in a reducing environment such as the cytoplasm, is somewhat unpredictable.
For this reason, a general and reliable method for the intracellular selection of Intrabodies was developed (Visintin et al, 1999). Based on the two-hybrid assay in yeast, the so-called “IACT” (Intracellular Antibody Capture Technology) allows us to isolate specific intrabodies, from diverse cDNA libraries of antibody domains [either ScFv (Single Chain Fragment Variable domains) or dAbs (single domain antibodies)] directly in the cytosolic environment, avoiding any protein manipulation and solving the folding problem.
File