Hinge influences in murine IgG binding to Cryptococcus neoformans capsule

Diane Sthefany Lima de Oliveira, Verenice Paredes, Adrielle Veloso Caixeta, Nicole Moreira Henriques, Maggie P. Wear, Patrícia Albuquerque, Maria Sueli Soares Felipe, Arturo Casadevall, André Moraes Nicola

Research output: Contribution to journalArticlepeer-review


Decades of studies on antibody structure led to the tenet that the V region binds antigens while the C region interacts with immune effectors. In some antibodies, however, the C region affects affinity and/or specificity for the antigen. One example is the 3E5 monoclonal murine IgG family, in which the mIgG3 isotype has different fine specificity to the Cryptococcus neoformans capsule polysaccharide than the other mIgG isotypes despite their identical variable sequences. Our group serendipitously found another pair of mIgG1/mIgG3 antibodies based on the 2H1 hybridoma to the C. neoformans capsule that recapitulated the differences observed with 3E5. In this work, we report the molecular basis of the constant domain effects on antigen binding using recombinant antibodies. As with 3E5, immunofluorescence experiments show a punctate pattern for 2H1-mIgG3 and an annular pattern for 2H1-mIgG1; these binding patterns have been associated with protective efficacy in murine cryptococcosis. Also as observed with 3E5, 2H1-mIgG3 bound on ELISA to both acetylated and non-acetylated capsular polysaccharide, whereas 2H1-mIgG1 only bound well to the acetylated form, consistent with differences in fine specificity. In engineering hybrid mIgG1/mIgG3 antibodies, we found that switching the 2H1-mIgG3 hinge for its mIgG1 counterpart changed the immunofluorescence pattern to annular, but a 2H1-mIgG1 antibody with an mIgG3 hinge still had an annular pattern. The hinge is thus necessary but not sufficient for these changes in binding to the antigen. This important role for the constant region in antigen binding could affect antibody biology and engineering.

Original languageEnglish (US)
Pages (from-to)110-121
Number of pages12
Issue number1
StatePublished - Jan 2022


  • Cryptococcus neoformans
  • antibody
  • capsule
  • immunofluorescence
  • isotype

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology


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