Abstract
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.
Highlights
Sharp and Dohme Research Laboratories, Davies, 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
Human rhinoviruses (HRVs)’ are picornaviruses that are responsible for most common colds in humans [1]
This notion was supported by human clinical studies showing that intranasally administered mAb lA6 was effective in modulating the symptoms of HRV infection [7]
Summary
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. We previously isolated a mouse monoclonal antibody, designated mAb lA6, that prevented HRV major group infection of HeLa cells by blocking virus attachment to cellular receptors [3]. Challenge with high titer HRV inocula failed to bypass the receptor blockade, suggesting that mAb lA6 or its derivatives may be useful in the prevention or treatment of common colds in humans This notion was supported by human clinical studies showing that intranasally administered mAb lA6 was effective in modulating the symptoms of HRV infection [7]. ’ The abbreviations used are: HRV, human rhinovirus; mAb, monoclonal antibody; IPTG, isopropylthiogalactoside; H, heavy; L, light; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; Ab, antibody; HPLC, high performance liquid chromatography
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