title: Agglutinating mouse IgG3 compares favourably with IgMs in typing of the blood group B antigen: Functionality and stability studies.
abstract: Mouse immunoglobulins M (IgMs) that recognize human blood group antigens induce haemagglutination and are used worldwide for diagnostic blood typing. Contrary to the current belief that IgGs are too small to simultaneously bind antigens on two different erythrocytes, we obtained agglutinating mouse IgG3 that recognized antigen B of the human ABO blood group system. Mouse IgG3 is an intriguing isotype that has the ability to form Fc-dependent oligomers. However, F(ab')2 fragments of the IgG3 were sufficient to agglutinate type B red blood cells; therefore, IgG3-triggered agglutination did not require oligomerization. Molecular modelling indicated that mouse IgG3 has a larger range of Fab arms than other mouse IgG subclasses and that the unique properties of mouse IgG3 are likely due to the structure of its hinge region. With a focus on applications in diagnostics, we compared the stability of IgG3 and two IgMs in formulated blood typing reagents using an accelerated storage approach and differential scanning calorimetry. IgG3 was much more stable than IgMs. Interestingly, the rapid decrease in IgM activity was caused by aggregation of the molecules and a previously unknown posttranslational proteolytic processing of the μ heavy chain. Our data point to mouse IgG3 as a potent diagnostic tool.
authors.affiliation: Malopolska Centre of Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7A, 30-387 Kraków, Poland.|Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387 Kraków, Poland.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Klaus
authors.affiliation: Malopolska Centre of Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7A, 30-387 Kraków, Poland.|Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387 Kraków, Poland.
authors.forename: Monika
authors.initials: M
authors.lastname: Bzowska
authors.affiliation: Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387 Kraków, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Kulesza
authors.affiliation: Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387 Kraków, Poland.
authors.forename: Agnieszka Martyna
authors.initials: AM
authors.lastname: Kabat
authors.affiliation: Malopolska Centre of Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7A, 30-387 Kraków, Poland.|Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387 Kraków, Poland.
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Jemioła-Rzemińska
authors.affiliation: Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387 Kraków, Poland.
authors.forename: Dominik
authors.initials: D
authors.lastname: Czaplicki
authors.affiliation: Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387 Kraków, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Makuch
authors.affiliation: Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387 Kraków, Poland.
authors.forename: Jarosław
authors.initials: J
authors.lastname: Jucha
authors.affiliation: Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387 Kraków, Poland.
authors.forename: Alicja
authors.initials: A
authors.lastname: Karabasz
authors.affiliation: Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University in Kraków, Gronostajowa 7, 30-387 Kraków, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Bereta
mesh_terms: D000017:ABO Blood-Group System / Q000097:blood*; D000818:Animals; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D006384:Hemagglutination; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry*; D007075:Immunoglobulin M / Q000737:chemistry*; D051379:Mice
chemical_list: D000017:ABO Blood-Group System; D007074:Immunoglobulin G; D007075:Immunoglobulin M