Preparation of integrin alpha(v)beta3-targeting Ab 38C2 constructs. Academic Article uri icon

Overview

abstract

  • This protocol describes the preparation of Ab constructs using agents that target cells expressing integrins alpha(v)beta3 and alpha(v)beta5, and the monoclonal aldolase Ab 38C2. The targeting agents are equipped with a diketone or vinylketone linker, and selectively react through the reactive Lys residues in the Ab binding sites to form 38C2 conjugates or chemically programmed 38C2 (i.e., cp38C2). The targeting agent possessing a diketone linker reacts with the Lys residues forming an enaminone derivative. By contrast, the vinylketone linker is used as the corresponding acetone adduct (i.e., a pro-vinylketone linker), and this pro-adapter undergoes a 38C2-catalyzed retro-aldol reaction to produce the vinylketone linker, which forms a Michael-type adduct with the Lys residues. The Ab construct formation is achieved in <1 h for the diketone compounds at ambient temperature, and in 2-16 h using the pro-vinylketone linker at 37 degrees C. The 38C2 constructs are retargeted to cells over-expressing integrins, and are potential candidates for immunotherapy.

publication date

  • January 1, 2007

Research

keywords

  • Antibodies, Catalytic
  • Drug Delivery Systems
  • Fructose-Bisphosphate Aldolase
  • Immunoglobulin Fab Fragments
  • Immunologic Techniques
  • Integrin alphaVbeta3

Identity

Scopus Document Identifier

  • 34347272702

Digital Object Identifier (DOI)

  • 10.1038/nprot.2007.3

PubMed ID

  • 17406606

Additional Document Info

volume

  • 2

issue

  • 2