Development of polyclonal and monoclonal antibodies for immunoassay and neutralization of human interleukin-4

J Immunol Methods. 1989 Feb 8;117(1):67-81. doi: 10.1016/0022-1759(89)90120-8.


A rabbit antiserum to partly purified recombinant E. coli-expressed human interleukin-4 (IL-4) has been produced which neutralizes the T cell growth factor, B cell growth factor, and Fc epsilon R2/CD23 inducing activities of IL-4. The antiserum demonstrated sufficient avidity to immunoprecipitate labelled COS7-expressed recombinant human IL-4. In contrast, rabbits immunized with conjugates of various synthetic IL-4 oligopeptides produced antisera which recognized IL-4 in both enzyme-linked immunosorbent assay (ELISA) and Western blotting formats, but failed to immunoprecipitate IL-4 from solution, or to neutralize bioactivity. Two rat monoclonal antibodies, 11B4, 22C10 were produced from a rat immunized with purified COS7 cell-expressed IL-4. These IgG2a antibodies recognized both E. coli-expressed and mammalian cell-expressed (COS7 and L cell) recombinant human IL-4 in solution (immunoprecipitation), as well as on solid phase (indirect ELISA and dot-blotting). The 11B4 antibody inhibited IL-4 bioactivity at an IC50 which was 25-50-fold in molar excess of factor. Both antibodies also recognized IL-4 bound to an immobilized rabbit IgG fraction of anti-IL-4. The 11B4 antibody was used to develop an immunoenzymetric assay capable of detecting less than 100 pg of analyte/ml. Supernatants from PBL, activated under varying conditions were tested for IL-4 levels. PHA and ConA were found to induce a relatively low degree of IL-4 production by these PBL. An approximately ten-fold greater level of IL-4 production was observed when they were stimulated with A23187 in combination with PMA. Various patient sera and cell line supernatants were also tested. These IL-4 immunoreagents are important tools for further studies of IL-4 immunobiology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / analysis
  • Antibodies, Monoclonal / biosynthesis*
  • Humans
  • Immune Sera* / analysis
  • Immune Sera* / pharmacology
  • Immunoenzyme Techniques*
  • Interleukin-4
  • Interleukins / chemical synthesis
  • Interleukins / immunology*
  • Interleukins / isolation & purification
  • Male
  • Molecular Sequence Data
  • Neutralization Tests*
  • Rabbits
  • Rats
  • Rats, Inbred Lew
  • Recombinant Proteins / chemical synthesis
  • Recombinant Proteins / immunology
  • Recombinant Proteins / isolation & purification


  • Antibodies, Monoclonal
  • Immune Sera
  • Interleukins
  • Recombinant Proteins
  • Interleukin-4