Prevention of Human Rhinovirus Infection by Multivalent Fab Molecules Directed against ICAM-1

ABSTRACT We have developed a technology for improving avidity by making bivalent, trivalent, or tetravalent recombinant polypeptides. We designed tripartite proteins consisting of the Fab fragment of an antibody fused with a hinge derived from human immunoglobulin D that was further linked to polymerization domains derived from human coiled-coil proteins. We report here on the application of this method with a Fab domain directed against the major human rhinovirus receptor, intercellular adhesion molecule 1 (ICAM-1). Multivalent anti-ICAM-1 molecules were produced in bacteria and purified as soluble preassembled homogeneous proteins at high yield. These proteins successfully blocked rhinovirus infection in vitro, with the efficiency increasing from monomer to dimer, trimer, and tetramer. The diminished dissociation rate of these multivalent antibodies and their improved efficacy in preventing rhinovirus infection provide a foundation for producing prophylactic and therapeutic molecules against human rhinovirus, the causative agent of the majority of common colds.

[1]  D M Crothers,et al.  The influence of polyvalency on the binding properties of antibodies. , 1972, Immunochemistry.

[2]  T. Arakawa,et al.  Size-exclusion chromatography with on-line light-scattering, absorbance, and refractive index detectors for studying proteins and their interactions. , 1996, Analytical biochemistry.

[3]  F. Hayden,et al.  Chemotherapy of rhinovirus colds , 1988, Antimicrobial Agents and Chemotherapy.

[4]  D. Stein,et al.  Construction and characterization of a new shuttle vector, pLES2, capable of functioning in Escherichia coli and Neisseria gonorrhoeae. , 1983, Gene.

[5]  L. Hood,et al.  The nucleotide sequence of a human immnnoglobulin C-gamma-1 gene , 1982 .

[6]  J. Whelan,et al.  ATF-a0, a novel variant of the ATF/CREB transcription factor family, forms a dominant transcription inhibitor in ATF-a heterodimers. , 1994, The Journal of biological chemistry.

[7]  Leroy E. Hood,et al.  The nucleotide sequence of a human immunoglobulin C gamma1 gene , 1982, Nucleic Acids Res..

[8]  D. Staunton,et al.  A cell adhesion molecule, ICAM-1, is the major surface receptor for rhinoviruses , 1989, Cell.

[9]  Brad Snedecor,et al.  High Level Escherichia coli Expression and Production of a Bivalent Humanized Antibody Fragment , 1992, Bio/Technology.

[10]  E. Padlan,et al.  Anatomy of the antibody molecule. , 1994, Molecular immunology.

[11]  F. Hayden,et al.  Modification of experimental rhinovirus colds by receptor blockade. , 1988, Antiviral research.

[12]  T. Laue,et al.  Modern Analytical Ultracentrifugation , 1994, Emerging Biochemical and Biophysical Techniques.

[13]  N. Nomura,et al.  Isolation and characterization of a novel member of the gene family encoding the cAMP response element-binding protein CRE-BP1. , 1993, The Journal of biological chemistry.

[14]  F. Grosse,et al.  A cloning cartridge of lambda t(o) terminator. , 1987, Nucleic acids research.

[15]  P. S. Kim,et al.  A switch between two-, three-, and four-stranded coiled coils in GCN4 leucine zipper mutants. , 1993, Science.

[16]  J. Philo.  An improved function for fitting sedimentation velocity data for low-molecular-weight solutes. , 1997, Biophysical journal.

[17]  H. Heyneker,et al.  Nucleotide sequence of the gene for heat-stable enterotoxin II of Escherichia coli , 1983, Infection and immunity.

[18]  P. Schuck,et al.  Size-distribution analysis of macromolecules by sedimentation velocity ultracentrifugation and lamm equation modeling. , 2000, Biophysical journal.

[19]  N. Hilschmann,et al.  [The primary structure of a crystalline monoclonal immunoglobulin kappa-type L-chain, subgroup I (Bence-Jones protein Rei.); isolation and characterization of the tryptic peptides; the complete amino acid sequence of the protein; a contribution to the elucidation of the three-dimensional structure o , 1975, Hoppe-Seyler's Zeitschrift fur physiologische Chemie.

[20]  A. Plückthun,et al.  New protein engineering approaches to multivalent and bispecific antibody fragments. , 1997, Immunotechnology : an international journal of immunological engineering.

[21]  N. Guex,et al.  SWISS‐MODEL and the Swiss‐Pdb Viewer: An environment for comparative protein modeling , 1997, Electrophoresis.

[22]  R. Webster,et al.  Encyclopedia of virology , 1999 .

[23]  F. Grosse,et al.  A cloning cartridge of λ t0 terminator , 1987 .

[24]  J. Vieira,et al.  The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. , 1982, Gene.

[25]  Leroy Hood,et al.  Nucleotide Sequence of a Human Immunoglobulin Cγ4 Gene , 1981 .

[26]  M. Little,et al.  Affinity enhancement of a recombinant antibody: formation of complexes with multiple valency by a single-chain Fv fragment-core streptavidin fusion. , 1996, Protein engineering.

[27]  Y. Kurosawa,et al.  Development of an artificial antibody system with multiple valency using an Fv fragment fused to a fragment of protein A. , 1993, The Journal of biological chemistry.

[28]  J. Casasnovas,et al.  Kinetics and Thermodynamics of Virus Binding to Receptor. , 1995, The Journal of Biological Chemistry.