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      Production and Characterization of Human Monoclonal Antibody Fab Fragments to Vaccinia Virus from a Phage-Display Combinatorial Library

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          Abstract

          A combinatorial, phage-display library of human Fab antibody fragments was generated from IgG heavy chain (HC) and light chain (LC) genes cloned from the lymphocytes of a vaccinia virus (VACV)-immune donor. To ascertain the complexity of the library, nucleotide sequences of the variable regions of the HC and LC genes were determined. Fourteen distinct HC and 18 distinct LC (7 κ and 11 λ) that formed a combinatorial library of 22 Fabs were identified. Immune-precipitation of radiolabeled VACV revealed that at least six different VACV proteins were recognized by the antibodies. Plaque-reduction neutralization demonstrated that six of the Fabs neutralized VACV in the presence of anti-human antibody. ELISA studies indicated that 15 of the Fabs were cross-reactive with monkeypox virus.

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          Most cited references33

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          Antigenic subunits of Hantaan virus expressed by baculovirus and vaccinia virus recombinants.

          Baculovirus and vaccinia virus vectors were used to express the small (S) and medium (M) genome segments of Hantaan virus. Expression of the complete S or M segments yielded proteins electrophoretically indistinguishable from Hantaan virus nucleocapsid protein or envelope glycoproteins (G1 and G2), and expression of portions of the M segment, encoding either G1 or G2 alone, similarly yielded proteins which closely resembled authentic Hantaan virus proteins. The expressed envelope proteins retained all antigenic sites defined by a panel of monoclonal antibodies to Hantaan virus G1 and G2 and elicited antibodies in animals which reacted with authentic viral proteins. A Hantaan virus infectivity challenge model in hamsters was used to assay induction of protective immunity by the recombinant-expressed proteins. Recombinants expressing both G1 and G2 induced higher titer antibody responses than those expressing only G1 or G2 and protected most animals from infection with Hantaan virus. Baculovirus recombinants expressing only nucleocapsid protein also appeared to protect some animals from challenge. Passively transferred neutralizing monoclonal antibodies similarly prevented infection, suggesting that an antibody response alone is sufficient for immunity to Hantaan virus.
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            Poxvirus dilemmas--monkeypox, smallpox, and biologic terrorism.

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              Assembly of combinatorial antibody libraries on phage surfaces: the gene III site.

              A phagemid system was developed for the monovalent display of combinatorial antibody Fab libraries on the surface of filamentous phage M13. Fab fragments were fused to the carboxyl-terminal domain of the gene III protein. Phage displaying Fab fragments on their surface, or Phabs, were enriched by 10(3)- to 10(5)-fold on antigen-coated surfaces over nonspecific phage. The method may replace current antibody cloning techniques.
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                Author and article information

                Journal
                Virology
                Virology
                Virology
                Published by Elsevier Inc.
                0042-6822
                1096-0341
                25 May 2002
                25 May 1999
                25 May 2002
                : 258
                : 1
                : 189-200
                Affiliations
                [a ]Virology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland, 21702-5011
                [b ]United States Army Medical Research Institute of Infectious Diseases, Geo-Centers, Inc. Fort Detrick, Frederick, Maryland, 21702-5011
                Author notes
                [1]

                To whom reprint requests should be addressed. Fax: (301) 619-2439. E-mail:Cschmalj@detrick.army.mil.

                Article
                S0042-6822(99)99701-5
                10.1006/viro.1999.9701
                9631611
                10329580
                f222c604-3694-48e5-aafe-78ff729da657
                Copyright © 1999 Published by Elsevier Inc.

                Elsevier has created a Monkeypox Information Center (https://www.elsevier.com/connect/monkeypox-information-center) in response to the declared public health emergency of international concern, with free information in English on the monkeypox virus. The Monkeypox Information Center is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its monkeypox related research that is available on the Monkeypox Information Center - including this research content - immediately available in publicly funded repositories, with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the Monkeypox Information Center remains active.

                History
                : 26 January 1999
                : 18 February 1999
                : 10 March 1999
                Categories
                Regular Article

                Microbiology & Virology
                Microbiology & Virology

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