Bacterial expression of antibody fragments that block human rhinovirus infection of cultured cells.
Human rhinoviruses are the major causative agents of the common cold in humans and have been divided into major and minor groups based on receptor specificity. cDNAs encoding the light and heavy chains of a murine monoclonal antibody that recognizes the major group receptor were cloned, abundantly expressed in Escherichia coli, and renatured into Fab fragments that blocked virus binding and protected HeLa cell monolayers from rhinovirus infection. Elimination of the cysteines normally bridging the heavy and light chains yielded molecules indistinguishable from wild-type Fab fragments in virus binding assays. Single-chain antibodies with covalently linked light and heavy variable domains were also expressed and showed receptor binding and cell protection activities. These recombinant antibody fragments are potentially useful in preventing or treating common colds in humans.
Condra
J H
JH
Department of Virus and Cell Biology, Merck Sharp and Dohme Research Laboratories, West Point, Pennsylvania 19486.
Sardana
V V
VV
Tomassini
J E
JE
Schlabach
A J
AJ
Davies
M E
ME
Lineberger
D W
DW
Graham
D J
DJ
Gotlib
L
L
Colonno
R J
RJ
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D002472:Cell Transformation, Viral; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005803:Genes, Immunoglobulin; D006367:HeLa Cells / Q000502:physiology; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011991:Receptors, Virus / Q000276:immunology / Q000502:physiology*; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D012229:Rhinovirus / Q000235:genetics / Q000276:immunology* / Q000502:physiology
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D011991:Receptors, Virus; D011994:Recombinant Proteins