Table IV.

Binding of monoclonal healthy immunized donor-derived RFs to chimeric IgG containing mutations in CH3 at positions 435-436: reactivity with the IgG3 polymorphism at 436a

Binding PatternRF:Chimeric IgG:Comments
3601 4-4-4-3 His-Tyr3602 4-4-4-3 His-Phe3603 3-3-3-3 His-Tyr3604 3-3-3-3 His-Phe3605 4-4-4-4 Arg-Tyr3611 3-3-3-3 Arg-Phe
GaMR-186.7 ± 2.878.1 ± 4.56.9 ± 0.252.2 ± 3.699.8 ± 3.014.8 ± 1.2Phe436 contributes
GaMR-2b84.5 ± 7.642.1 ± 1.02.5 ± 1.019.1 ± 0.610.9 ± 9.418.4 ± 4.7
GaMR-393.7 ± 8.637.9 ± 0.72.6 ± 0.230.5 ± 1.192.9 ± 4.117.7 ± 0.4
GaMR-574.0 ± 9.161.4 ± 4.911.6 ± 0.220.1 ± 3.161.6 ± 8.916.4 ± 0.4
GaMR-1287.3 ± 4.172.6 ± 1.79.6 ± 1.718.8 ± 1.166.5 ± 9.415.5 ± 1.5
GaMR-13b73.7 ± 3.677.6 ± 1.08.8 ± 4.529.0 ± 3.559.3 ± 7.120.0 ± 0.3
GaMR-14b65.4 ± 4.479.4 ± 2.910.1 ± 2.919.6 ± 2.290.0 ± 6.316.1 ± 0.2
GaMR-2096.2 ± 3.566.3 ± 4.013.9 ± 0.525.6 ± 4.991.8 ± 3.124.1 ± 1.9
GaMR-25b73.1 ± 6.248.2 ± 2.111.6 ± 5.523.1 ± 0.979.3 ± 2.119.1 ± 1.5
GaMR-2787.8 ± 4.948.8 ± 0.98.3 ± 3.321.2 ± 0.46.3 ± 5.126.7 ± 0.1
GaMR-28b78.9 ± 2.362.2 ± 3.66.8 ± 1.168.1 ± 1.388.0 ± 1.061.4 ± 2.2Anti-allotype, Phe436 IgG3
GaMR-30b74.3 ± 4.585.8 ± 0.36.7 ± 0.723.1 ± 1.299.8 ± 1.022.4 ± 0.1
GaMR-33b74.8 ± 9.691.0 ± 1.411.2 ± 3.329.8 ± 6.592.1 ± 1.418.3 ± 1.9
GaMR-37b74.1 ± 9.082.5 ± 3.910.5 ± 3.229.6 ± 3.693.8 ± 3.919.3 ± 0.3
GaMR-3997.1 ± 8.535.5 ± 0.17.6 ± 2.019.2 ± 0.66.5 ± 0.617.0 ± 2.9
GaMR-4168.7 ± 1347.4 ± 1.47.9 ± 0.716.4 ± 0.37.7 ± 1.215.2 ± 0.7
GaD1-278.0 ± 2.141.0 ± 1.310.5 ± 2.717.1 ± 3.412.0 ± 7.59.6 ± 1.9
GaFO-373.3 ± 2.792.0 ± 2.173.6 ± 2.2108.4 ± 6.842.4 ± 2.410.8 ± 1.8
GaTT-386.9 ± 4.946.5 ± 2.08.5 ± 1.943.3 ± 1.58.6 ± 1.252.0 ± 1.1Anti-allotype, Phe436 IgG3
GaTT-780.9 ± 4.577.6 ± 4.010.3 ± 1.131.6 ± 0.96.8 ± 0.941.2 ± 2.6Anti-allotype, Phe436 IgG3
GaTT-951.0 ± 3.783.5 ± 1.64.3 ± 0.790.4 ± 3.292.7 ± 1.1100.4 ± 0.6Anti-allotype, Phe436 IgG3
PanMR-2477.5 ± 0.460.0 ± 1.267.2 ± 0.582.7 ± 1.444.0 ± 0.7100.7 ± 0.5
1, 3, 4D1-486.0 ± 0.724.0 ± 0.693.7 ± 1.49.0 ± 0.3103.1 ± 1.497.1 ± 1.1Tyr436 important
1, 2, 3LN-1174.9 ± 1.384.3 ± 3.549.3 ± 5.142.8 ± 0.924.7 ± 1.2102.8 ± 0.6
1, 4MR-16b77.4 ± 1.011.2 ± 1.47.5 ± 3.717.6 ± 1.55.0 ± 2.016.1 ± 0.8Tyr436 important
  • a Site-directed mutations at positions 435 and/or 436 in the CH3 of IgG3 or IgG4 were introduced in the CH3 domain of IgG3 or IgG4 as represented schematically at the top of each column. Mutated CH3 domains at these positions were also shuffled into hybrid proteins generated from the IgG3 and IgG4 genes. The number 3 or 4 indicates whether the exon originated from the IgG3 or IgG4 heavy chain gene. Results are the mean of six determinations from two separate experiments ± SEM for each RF expressed as a percentage binding to IgG4 defined as 100%. For RFs DI-4 and LN-11, the results are expressed as a percentage ± SEM of binding to IgG3 defined as 100%.

  • b Clonally related RFs.