Article Version 1 Preserved in Portico This version is not peer-reviewed
Version 1 : Received: 1 November 2018 / Approved: 2 November 2018 / Online: 2 November 2018 (10:42:20 CET)
How to cite: Yang, D.; Kroe-Barrett, R.; Singh, S.; Laue, T. IgG Charge. Preprints 2018, 2018110052. https://doi.org/10.20944/preprints201811.0052.v1. Yang, D.; Kroe-Barrett, R.; Singh, S.; Laue, T. IgG Charge. Preprints 2018, 2018110052. https://doi.org/10.20944/preprints201811.0052.v1.
It has been known since the 1930’s that all immunoglobulins carry a weak negative charge in physiological solvents. However, there has been no systematic exploration of this fundamental property. Accurate charge measurements have been made using membrane confined electrophoresis in two solvents (pH 5.0 and pH 7.4) on a panel of twelve mAb IgGs, as well as their F(ab’)2 and Fc fragments. The following observations were made at pH 5.0: 1) the measured charge differs from the calculated charge by ~40 for the intact IgGs, and by ~20 for the Fcs; 2) the intact IgG charge depends on both Fv and Fc sequences, but does not equal the sum of the F(ab)’2 and Fc charge; 3) the Fc charge is consistent within a class. In phosphate buffered saline, pH 7.4: 1) the intact IgG charges ranged from 0 to -13; 2) the F(ab’)2 fragments are nearly neutral for IgG1s and IgG2s, and about -5 for some of the IgG4s; 3) all Fc fragments are weakly anionic, with IgG1 < IgG2 < IgG4; 4) the charge on the intact IgGs does not equal the sum of the F(ab’)2 and Fc charge. In no case is the calculated charge, based on H+ binding, remotely close to the measured charge. The charge on IgGs in physiological solvent is sufficiently small to minimize its contribution to thermodynamic nonideality. Some of the mAbs carried a charge in physiological salt that was outside the range observed for serum-purified human poly IgG. To best match physiological properties, a therapeutic mAb should have a measured charge that falls within the range observed for serum-derived human IgGs.
analytical electrophoresis; IgG subclasses; monoclonal IgG; protein charge
LIFE SCIENCES, Biochemistry
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