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
Date Published1994 Sep 2
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

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.

Alternate JournalJ. Mol. Biol.
PubMed ID8071997