Generation and selection of monoclonal antibodies, single-chain Fv and antibody fusion phage specific for human melanoma-associated antigens

Most routine testing for plant viruses is currently carried out using monoclonal and polyclonal antibodies. Traditional methods of antibody production however can be time consuming and require the use of expensive cell culture facilities. Recombinant antibody technology however is starting to make an impact in this area, enabling the selection of antibody fragments in a few weeks compared with the many months associated with traditional methods and requires only basic microbiological facilities. Single chain Fv antibody fragments (scFv) have been selected from a synthetic phage-antibody library by affinity selection with purifiedPotato virus Y, ordinary strain (PVYO). The scFv selected was specific for PVY and detected 7 out of 9 isolates of PVYOwhilst it did not detect 15 isolates from the closely related necrotic strains PVYNand PVYNTN. In ELISA the scFv could be used to detect virus at concentrations of 50 ng/ml in plant sap and was shown to have similar limits of detection as commercially available PVY monoclonal antibodies. These results highlight the potential of the technology for the selection of strain specific antibodies with an affinity and assay sensitivity similar to traditional monoclonal antibodies and their use in viral diagnostics.

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Identification of Residues That Stabilize the Single-chain Fv of Monoclonal Antibodies B3

Journal of Biological Chemistry ◽

10.1074/jbc.270.40.23373 ◽

1995 ◽

Vol 270 (40) ◽

pp. 23373-23380 ◽

Cited By ~ 25

Author(s):

Itai Benhar ◽

Ira Pastan

Keyword(s):

Monoclonal Antibodies ◽

Single Chain ◽

Single Chain Fv

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Selection of human antitumor single-chain Fv antibodies from the B-cell repertoire of patients immunized against autologous neuroblastoma

Medical and Pediatric Oncology ◽

10.1002/1096-911x(20001201)35:6<692::aid-mpo45>3.0.co;2-7 ◽

2000 ◽

Vol 35 (6) ◽

pp. 692-695 ◽

Cited By ~ 4

Author(s):

Claudia Rossig ◽

Jed G. Nuchtern ◽

Malcolm K. Brenner

Keyword(s):

B Cell ◽

Single Chain ◽

Cell Repertoire ◽

B Cell Repertoire ◽

Single Chain Fv ◽

Selection Of

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Generation and Selection of a Panel of Pan-Filovirus Single-Chain Antibodies using Cell-Free Ribosome Display

10.1101/327296 ◽

2018 ◽

Author(s):

Adinarayana Kunamneni ◽

Elizabeth C. Clarke ◽

Chunyan Ye ◽

Steven B. Bradfute ◽

Ravi Durvasula

Keyword(s):

Hemorrhagic Fever ◽

Cross Reactivity ◽

Free Ribosome ◽

Rapid Diagnostics ◽

Single Chain ◽

Ribosome Display ◽

Single Chain Antibodies ◽

Single Chain Fv ◽

New Generation ◽

Selection Of

AbstractFiloviruses, which include ebolaviruses and marburgvirus, can cause outbreaks of highly lethal hemorrhagic fever. This disease causes significant morbidity and mortality in humans and non-human primates, with human fatality rates reaching 90% during some outbreaks. Currently, there are a lack of licensed vaccines or antivirals for these viruses. Since early symptoms of filovirus infection mimic more common diseases, there is a strong unmet public health and biodefense need for broad-spectrum filovirus rapid diagnostics. We have generated a panel of mouse single-chain Fv-antibodies (scFvs) to filovirus glycoproteins (GPs) using cell-free ribosome display and determined their cross-reactivity profiles to all known filovirus species. Two scFvs (4-2 and 22-1) were able to detect all known Ebolavirus and Marburgvirus species. This is the first report on ribosome display scFvs that can detect a broad set of filovirus GPs, which demonstrates their potential use in the development of a new generation of rapid diagnostic immunoassays.

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