Part:BBa_K3929015

Gene_Nb_CDR1_5773

Function: Gene sequence expressing nanobody CDR1.

Usage and biology.

Antibodies are daily use and have multiple applications in biomedical sciences because of their versatility and high affinity. This is the reason why nanobodies, a single domain antibodies derived from camelid, has been created. They have the same function as antibodies, but they are smaller and highly stable. Their useful size enable analysis of complex macromolecules and small proteins.

Fragments of antibodies are widely used in technology and medicine. The arms are cut off, forming smaller molecules known as Fabs. Each Fab contains a single site for binding to their target molecules. If this Fab binds to a cell surface protein, it can be used as a diagnostic tool to identify damaged or malignant cells. If this Fab is connected to a fluorescent molecule, it can be used to image the location of its target molecule inside cells. If the Fab binds to a viral protein, it can be used as a drug to fight the virus. Unfortunately, Fab molecules are rather large, and since they are composed of two chains, they are not as stable as we would like for these applications. This is where nanobodies come in (1).

Nanobody (Nb) sequence has been obtained from a llama nanobody that already exists and works. It has been divided into three sequences: CDR1, CDR2 and CDR3 to mix them with other CDR sequences from different nanobodies from different species of camelids. It will be used for nanobody gene assembly by in vitro recombination. Each CDR was chemically synthesized by IDT.

Risk group: 1 (2). Disease risk to humans: no. Risk to environment: no.

References: 1.David S. Goodsell, Christine Zardecki, Helen M. Berman, Stephen K. Burley. (2020) Insights from 20 years of the Molecule of the Month. BAMBed 48: 350-355. 2.National Institute of Occupational Health and Safety (INSHT) about exposure to biological agents (REAL DECRETO 664/1997).

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 7
  Illegal PstI site found at 29
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 7
  Illegal PstI site found at 29
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 7
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 7
  Illegal PstI site found at 29
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 7
  Illegal PstI site found at 29
  
• 1000
  COMPATIBLE WITH RFC[1000]