The immunogenicity of the tumor-associated antigen Lewis(y) may be suppressed by a bifunctional cross-linker required for coupling to a carrier protein

Chemistry. 2004 Jul 19;10(14):3517-24. doi: 10.1002/chem.200400074.

Abstract

A Lewis(y) (Le(y)) tetrasaccharide modified by an artificial aminopropyl spacer was synthesized by a highly convergent approach that employed a levulinoyl ester and a 9-fluorenylmethoxycarbonate for temporary protection of the hydroxy groups and a trichloroethyloxycarbonyl as an amino protecting group. The artificial aminopropyl moiety was modified by a thioacetyl group, which allowed efficient conjugation to keyhole limpet hemocyanin (KLH) modified by electrophilic 4-(maleimidomethyl)cyclohexane-1-carboxylate (MI). Mice were immunized with the KLH-MI-Le(y) antigen. A detailed analysis of sera by ELISA established that a strong immunoglobulin G (IgG) antibody response was elicited against the linker region. The use of a smaller and more flexible 3-(bromoacetamido)propionate for the attachment of Le(y) to KLH not only reduced the IgG antibody response against the linker but also led to a significantly improved immune response against the Le(y) antigen. This study shows that highly antigenic linkers suppress antibody responses to weak antigens such as self-antigens.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antibodies / immunology
  • Carbohydrate Conformation
  • Carrier Proteins / chemistry*
  • Carrier Proteins / metabolism*
  • Cross-Linking Reagents / chemistry*
  • Enzyme-Linked Immunosorbent Assay
  • Hydrolysis
  • Immunization
  • Immunogenetics
  • Lewis Blood Group Antigens / chemistry*
  • Lewis Blood Group Antigens / genetics
  • Lewis Blood Group Antigens / immunology*
  • Maleimides / chemistry
  • Neoplasms / genetics*
  • Neoplasms / immunology*
  • Polysaccharides / chemistry
  • Polysaccharides / genetics
  • Polysaccharides / immunology
  • Protein Binding

Substances

  • Antibodies
  • Carrier Proteins
  • Cross-Linking Reagents
  • Lewis Blood Group Antigens
  • Lewis Y antigen
  • Maleimides
  • Polysaccharides
  • maleimide