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

GOLDBAUM F.A.; FIELDS B.A.; CAUERFF A.; YSERN X.; HOUDUSSE A.; EISELE JL; POLJAK R.J.; MARIUZZA R.A.

JOURNAL OF MOLECULAR BIOLOGY

Elsevier

Año: 1994 p. 739 - 743

0022-2836

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.