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Crystallization of intact monoclonal antibodies

Published on Oct 1, 1995in Proteins2.501
· DOI :10.1002/prot.340230218
Lisa J. Harris3
Estimated H-index: 3
(UCR: University of California, Riverside),
Eileen Skaletsky3
Estimated H-index: 3
,
Alexander McPherson51
Estimated H-index: 51
(UCR: University of California, Riverside)
Abstract
Attempts were made to crystallize four monoclonal antibodies, one IgG2ak and three IgG1k. Using a PEG 3350 screen combined with detergents, and developed from our experiments with an IgG2ak antibody specific for canine lymphoma cells,1,2 crystals have now been obtained of two of these four immunoglobulins, an antiphenytoin and an antiphenobarbital antibody. A complex between the antiphenobarbital antibody and its drug antigen crystallized as well. The antibody for phenytoin has, to this point, produced only clustered microcrystals, marginally suitable for X-ray analysis. Single crystals of the IgG1k antibody against phenobarbital, however, were characterized by X-ray diffraction to be primitive monoclinic, with unit cell dimensions a = 67 A, b = 193 A, c = 74 A, and β = 110°. These crystals have an entire IgG1k molecule as the asymmetric unit and they diffract to at least 3.2 A resolution. © 1995 Wiley-Liss, Inc.
  • References (17)
  • Citations (50)
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References17
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Abstract The structures of the various regions of an antibody molecule are analysed and correlated with biological function. The structural features which relate to potential applications are detailed.
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Conformational changes in immunoglobulin fragments on complexation were first noted in trigonal crystals (ammonium sulphate form) of the Meg light chain dimer in 1974 (Edmundson et al., 1974). Additional studies with Meg crystals demonstrated how induced fit mechanisms increase the diversity of ligands which can be bound by a single active site (Ely et al., 1978; Edmundson el al., 1984, 1987, 1989, 1993; Edmundson and Ely, 1985). The ligandinduced conformational changes in the Meg dimer were all...
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#1Steven Sheriff (BMS: Bristol-Myers Squibb)H-Index: 28
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