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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\n                    <jats:italic>Cryptococcus neoformans<\/jats:italic>\n                    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\n                    <jats:italic>C<\/jats:italic>\n                    .\u00a0\n                    <jats:italic>neoformans<\/jats:italic>\n                    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\u2010mIgG3 and an annular pattern for 2H1\u2010mIgG1; these binding patterns have been associated with protective efficacy in murine cryptococcosis. Also as observed with 3E5, 2H1\u2010mIgG3 bound on ELISA to both acetylated and non\u2010acetylated capsular polysaccharide, whereas 2H1\u2010mIgG1 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\u2010mIgG3 hinge for its mIgG1 counterpart changed the immunofluorescence pattern to annular, but a 2H1\u2010mIgG1 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.\n                  <\/jats:p>","DOI":"10.1111\/imm.13411","type":"journal-article","created":{"date-parts":[[2021,8,30]],"date-time":"2021-08-30T04:39:13Z","timestamp":1630298353000},"page":"110-121","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Hinge influences in murine IgG binding to\n                    <i>Cryptococcus neoformans<\/i>\n                    capsule"],"prefix":"10.1111","volume":"165","author":[{"given":"Diane Sthefany Lima de","family":"Oliveira","sequence":"first","affiliation":[{"name":"Institute of Biological Sciences University of Bras\u00edlia  Bras\u00edlia Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Verenice","family":"Paredes","sequence":"additional","affiliation":[{"name":"Faculty of Medicine University of Bras\u00edlia  Bras\u00edlia Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Adrielle Veloso","family":"Caixeta","sequence":"additional","affiliation":[{"name":"Faculty of Medicine University of Bras\u00edlia  Bras\u00edlia Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicole Moreira","family":"Henriques","sequence":"additional","affiliation":[{"name":"Unieuro University Center  Bras\u00edlia Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maggie P.","family":"Wear","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology &amp; Immunology Johns Hopkins Bloomberg School of Public Health  Baltimore MD USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Patr\u00edcia","family":"Albuquerque","sequence":"additional","affiliation":[{"name":"Faculty of Ceil\u00e2ndia University of Bras\u00edlia  Bras\u00edlia Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maria Sueli Soares","family":"Felipe","sequence":"additional","affiliation":[{"name":"Graduate Program in Genomic Sciences and Biotechnology Catholic University of Bras\u00edlia  Bras\u00edlia Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arturo","family":"Casadevall","sequence":"additional","affiliation":[{"name":"Department of Molecular Microbiology &amp; Immunology Johns Hopkins Bloomberg School of Public Health  Baltimore MD USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8656-5835","authenticated-orcid":false,"given":"Andr\u00e9 Moraes","family":"Nicola","sequence":"additional","affiliation":[{"name":"Faculty of Medicine University of Bras\u00edlia  Bras\u00edlia Brazil"},{"name":"Graduate Program in Genomic Sciences and Biotechnology Catholic University of Bras\u00edlia  Bras\u00edlia Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2021,9,7]]},"reference":[{"key":"e_1_2_9_2_1","first-page":"1317","article-title":"The mechanism of diphtheria immunity and tetanus immunity in animals. 1890","volume":"28","author":"Behring E","year":"1991","journal-title":"Mol Immunol"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj0660677"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1042\/bj0660600"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00845a045"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.54.3.864"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0167-5699(92)90093-M"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.121.1.171"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.97.1.75"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2012-01-380121"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0065-2776(08)60019-X"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/nri2206"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.3389\/fmicb.2016.00022"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1209750109"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M700661200"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M806473200"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.169.3.1379"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.174.4.2132"},{"key":"e_1_2_9_19_1","doi-asserted-by":"crossref","first-page":"2659","DOI":"10.4049\/jimmunol.146.8.2659","article-title":"H chain C domains influence the strength of binding of IgG for streptococcal group A carbohydrate","volume":"146","author":"Cooper LJ","year":"1991","journal-title":"J Immunol"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/0167-5699(92)90120-V"},{"key":"e_1_2_9_21_1","doi-asserted-by":"crossref","first-page":"2231","DOI":"10.4049\/jimmunol.150.6.2231","article-title":"Role of heavy chain constant domains in antibody\u2010antigen interaction. 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