Preparation of MUC-1 oligomers using an improved convergent solid-phase peptide synthesis.

The Journal of Biological Chemistry
E KrambovitisK Barlos

Abstract

The sequentially repeating nature of the core mucin polypeptide chain MUC-1 on the surface of malignant cells makes it an excellent target for cancer immunotherapy. We describe a reliable and efficient method of synthesizing oligomers, up to five tandem repeats and oligomer heterotope derivatives with a 15-amino acid epitope from tetanus toxin using an improved convergent solid-phase peptide synthesis. The different oligomers were easily distinguishable by reverse-phase high pressure liquid chromatography, but they were poorly fixed and migrated with the same migration rate, irrespective of size, in electrophoretic studies. In contrast, the oligomer heterotopes exhibited size-dependent electrophoretic behavior but in high pressure liquid chromatography chromatograms the different heterotopes were eluted simultaneously in two peaks representing the L- and D-enantiomers of the derivatives. The oligomer heterotopes were recognized as antigens in Western blotting with a murine monoclonal antibody against the epitope APDTR. In enzyme immunoassay studies with the same antibody an increasing reactivity was observed against the larger oligomers and confirmed by inhibition assays as the MUC-1 pentamer was the most efficient inhibitor. T...Continue Reading

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Citations

May 14, 1999·Journal of Peptide Science : an Official Publication of the European Peptide Society·C ChivaF Albericio
Jun 21, 2001·Biochemical and Biophysical Research Communications·A ZafiropoulosE Krambovitis
Nov 1, 2002·Biochemical and Biophysical Research Communications·Stavroula BaritakiElias Krambovitis
Jul 4, 2013·Proceedings of the National Academy of Sciences of the United States of America·Ting Wang, Samuel J Danishefsky
Mar 3, 2007·Biochemical and Biophysical Research Communications·Apostolos K RizosElias Krambovitis
Jul 2, 2005·The Journal of Arthroplasty·John J CallaghanSteve S Liu
Mar 12, 2017·Amino Acids·Anthony BougasGeorge P Dinos
Nov 9, 2004·Chemistry : a European Journal·Gaëlle-Anne CremerAgnès Delmas
Nov 3, 2007·Journal of Peptide Science : an Official Publication of the European Peptide Society·Zoltán BánócziFerenc Hudecz
Jan 1, 2009·Biopolymers·Petros A GalanakisGeorgios A Spyroulias

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