Crystallization and preliminary X-ray diffraction study of an idiotope-anti-idiotope Fv-Fv complex.

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TitleCrystallization and preliminary X-ray diffraction study of an idiotope-anti-idiotope Fv-Fv complex.
Publication TypeJournal Article
Year of Publication1994
AuthorsGoldbaum, FA, Fields, BA, Cauerhff, A, Ysern, X, Houdusse, A, Eisele, JL, Poljak, RJ, Mariuzza, RA
JournalJ Mol Biol
Volume241
Issue5
Pagination739-43
Date Published1994 Sep 2
ISSN0022-2836
KeywordsAmino Acid Sequence, Antibodies, Anti-Idiotypic, Antibodies, Monoclonal, Antigen-Antibody Complex, Cloning, Molecular, Conserved Sequence, Crystallization, Crystallography, X-Ray, Genes, Immunoglobulin, Immunoglobulin Fragments, Immunoglobulin Variable Region, Isoelectric Focusing, Molecular Sequence Data, Muramidase
Abstract

A complex between the Fv fragment of an anti-hen eggwhite lysozyme antibody (D1.3) and the Fv fragment of an antibody specific for an idiotypic determinant of D1.3 has been crystallized in a form suitable for X-ray diffraction analysis. Both Fv fragments were expressed in soluble form in Escherichia coli and purified by affinity chromatography; diffraction-quality crystals were only obtained following separation of each Fv into distinct isoelectric forms. The crystals belong to space group C2, have unit cell dimensions a = 152.8 A, b = 79.4 A, c = 51.5 A, beta = 100.2 degrees, and diffract to better than 2.2 A resolution. The solvent content of the crystals is approximately 60% (v/v) with one Fv-Fv complex in the asymmetric unit. The ability to readily express both components of an antigen-antibody system in bacteria will allow us to rigorously assess the energetic contribution of individual amino acids to complex formation through pairwise mutagenesis of interacting residues.

DOI10.1006/jmbi.1994.1549
Alternate JournalJ. Mol. Biol.
PubMed ID8071997