Preparation and Characterization of Trastuzumab Fab-Conjugated Liposomes (Immunoliposomes)

Methods Mol Biol. 2023:2622:197-205. doi: 10.1007/978-1-0716-2954-3_17.

Abstract

Immunoliposomes are made by conjugating antibodies or antibody fragments on liposome surfaces. Antibody fragments Fab', single-chain Fv fragments (scFv), or new constructs such as nanobodies are commonly used instead of whole IgGs for reduced risk immunogenicity. Here we described the preparation and characterization of immunoliposome-containing trastuzumab Fabs on the surface. The targeting ligand Fab-PEG-DSPE was synthesized by site-specific coupling between the C-terminal cysteine of the Fab and the maleimide group at the distal end of a DSPE-PEG. It was incorporated into preformed liposomes at 60 °C above the lipid bilayer phase transition temperature. The binding avidity of the immunoliposomes containing different Fab valencies was characterized using biolayer interferometry.

Keywords: Avidity; Immunoliposomes; PEG-DSPE; Trastuzumab Fab fragment.

MeSH terms

  • Immunoglobulin Fab Fragments / chemistry
  • Immunoglobulin Fragments*
  • Liposomes* / chemistry
  • Trastuzumab

Substances

  • Liposomes
  • Trastuzumab
  • Immunoglobulin Fragments
  • Immunoglobulin Fab Fragments