Acid-induced structural changes of a mouse IgG2a monoclonal antibody (MN12) studied by transient electric birefringence measurement.
Acid-induced structural changes of a mouse IgG2a monoclonal antibody (MN12) as indicated by Jiskoot et al. (Eur. J. Biochem. 201,223-232 (1991)) were studied by measuring the transient electric birefringence of MN12 in aqueous solution and in glycerol-water mixtures at different pH conditions. A multi-exponential analysis program, DISCRETE (Provencher,S.W., Biophys.J.16,27-41 (1976)), and a constrained inverse Laplace transform program, CONTIN (Provencher, S.W., Comp. Phys. Comm. 27, 213-227 (1982)) have been used to determine the number of exponentials needed to represent the data and their decay times. Measurement of the time-resolved electric field induced birefringence makes it possible to study rotational processes on a timescale from several tens of nanoseconds to microseconds. This enabled us to monitor the segmental flexibility and the rotational motion of single antibody molecules as well as the occurrence of aggregates. The results show an increase in hydrodynamic dimensions of MN12 upon lowering the pH from 6.6 to 2.7. Additionally, the original segmental flexibility, which could be monitored for the samples in glycerol-water mixtures, is altered at low pH. The results have been interpreted as swelling of MN12 followed by dimerization.
van Haeringen
B
B
Dept. of Biophysics, Free University, Amsterdam, The Netherlands.
Jiskoot
W
W
van Grondelle
R
R
Bloemendal
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001718:Birefringence; D004560:Electricity; D005990:Glycerol / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry*; D051379:Mice; D008956:Models, Chemical; D011487:Protein Conformation; D012984:Software; D014867:Water / Q000737:chemistry
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D014867:Water; D005990:Glycerol