Applied Biochemistry and Biotechnology

, Volume 169, Issue 1, pp 159–169

Recombinant Single-Chain Antibody with the Trojan Peptide Penetratin Positioned in the Linker Region Enables Cargo Transfer Across the Blood–Brain Barrier

  • Nives Škrlj
  • Gorazd Drevenšek
  • Samo Hudoklin
  • Rok Romih
  • Vladka Čurin Šerbec
  • Marko Dolinar
Article

DOI: 10.1007/s12010-012-9962-7

Cite this article as:
Škrlj, N., Drevenšek, G., Hudoklin, S. et al. Appl Biochem Biotechnol (2013) 169: 159. doi:10.1007/s12010-012-9962-7

Abstract

Delivery of therapeutic proteins into tissues and across the blood–brain barrier (BBB) is limited by the size and biochemical properties of the proteins. Efficient delivery across BBB is generally restricted to small, highly lipophilic molecules. However, in the last decades, several peptides that can pass cell membranes have been identified. It has been shown that these peptides are also capable of delivering large hydrophilic cargoes into cells and are therefore a powerful biological tool for transporting drugs across cell membranes and even into the brain. We designed and prepared a single-chain antibody fragment (scFvs), specific for the pathological form of the prion protein (PrPSc), where a cell-penetrating peptide (CPP) was used as a linker between the two variable domains of the scFv. The intravenously administered recombinant scFv-CPP was successfully targeted to and delivered into mouse brain cells. Our single-chain antibody fragments are of special interest in view of possible therapeutic reagents design not only for prion diseases but also for other neurodegenerative diseases.

Keywords

Antibody fragment Cell-penetrating peptides Brain delivery Prion Penetratin 

Abbreviations

ABTS

2,2′-Azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)

CNS

Central nervous system

CPP

Cell-penetrating peptides

DAPI

4′,6-Diamidino-2-phenylindole

Fv

Antibody variable domains (Vl + Vh)

HRP

Horseradish peroxidase

IMAC

Immobilized metal ion affinity chromatography

IPTG

Isopropyl β-d-1-thiogalactopyranoside

LLH

scFv in Vl-linker-Vh chain arrangement

PBS

Phosphate-buffered saline

PrP

Prion protein

PrPC

Cellular form of the PrP

PrPSc

Pathogenic form of the PrP

scFv

Antibody single-chain fragment

TBS

Tris-buffered saline

TMB

3,3′,5,5′-Tetramethylbenzidine

Vh

Variable domain of the heavy-chain of an antibody

Vl

Variable domain of the light chain of an antibody

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Nives Škrlj
    • 1
  • Gorazd Drevenšek
    • 2
  • Samo Hudoklin
    • 3
  • Rok Romih
    • 3
  • Vladka Čurin Šerbec
    • 1
    • 4
  • Marko Dolinar
    • 1
  1. 1.Biochemistry Chair, Faculty of Chemistry and Chemical TechnologyUniversity of LjubljanaLjubljanaSlovenia
  2. 2.Institute of Pharmacology and Experimental Toxicology, Faculty of MedicineUniversity of LjubljanaLjubljanaSlovenia
  3. 3.Institute of Cell Biology, Faculty of MedicineUniversity of LjubljanaLjubljanaSlovenia
  4. 4.Department for Production of Diagnostic Reagents and ResearchBlood Transfusion Centre of SloveniaLjubljanaSlovenia

Personalised recommendations