Clinical Chemistry
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Clinical Chemistry 20: 1071-1075, 1974;
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Clinical Chemistry, Vol 20, 1071-1075, Copyright © 1974 by the American Association for Clinical Chemistry

Kinetics of the IgG—anti-IgG Reaction, as Evaluated by Conventional and Stopped-Flow Nephelometry

John Savory 1, Gregory Buffone 1, and Richard Reich 1

1 Division of Clinical Chemistry, Departments of Medicine, Pathology, and Biochemistry, the University of North Carolina, Chapel Hill, N. C. 27514; and American Instrument Co., Silver Spring, Md. 20910.

A stopped-flow spectrophotofluorometer equipped with a data-acquisition system was used to study the rate of formation of IgG—anti-IgG complexes by nephelometry. Light-scattering aggregates could be detected within 100 ms. Early rates ( 0-3 s) increased with antigen concentration, while rates during later stages (5-100 s) followed the same trend exhibited by the precipitin curve. Polyethylene glycol (known to exert a profound effect on antigen—antibody reactions) at a concentration of 40 g/liter affected the IgG—anti-IgG reaction as follows: there was an initial lag phase of 5-10 s and a subsequent rapid reaction over the next 20-40 s, characterized by a four-to five-fold enhancement in the amount of light scattered. Our observations demonstrate that centrifugal-analyzer techniques can be used for the kinetic measurement of specific proteins in serum.


Key Words: spectrophotofluorometer • stopped-flow kinetic analysis for proteins • polyethylene glycol

Submitted on March 21, 1974
Accepted on May 24, 1974






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Copyright © 1974 by the American Association for Clinical Chemistry.