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Clinical and Diagnostic Laboratory Immunology, September 1998, p. 636-644, Vol. 5, No. 5
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Selection of Recombinant, Library-Derived Antibody Fragments against p24 for Human Immunodeficiency Virus Type 1 Diagnostics

Hans J. W. de Haard,1,* Bert Kazemier,1 Marck J. M. Koolen,1 Liekle J. Nijholt,1 Rob H. Meloen,2 Bob van Gemen,1 Hennie R. Hoogenboom,3 and Jan-Willem Arends3,4

Biosciences Research Unit, Organon Teknika, Boxtel,1 Department of Molecular Recognition, Institute for Animal Science and Health (ID-DLO), Lelystad,2 and CESAME, Department of Pathology, University Hospital,3 and Maastricht University,4 Maastricht, The Netherlands

Received 4 February 1998/Returned for modification 29 April 1998/Accepted 22 June 1998

By application of combinatorial library technology, we generated the first recombinant antibody fragments directed against the major capsid protein p24 of human immunodeficiency virus type 1 (HIV-1). A library of single-chain Fv fragments (scFvs) was constructed by using the antibody variable-region (V) genes of B cells derived from the spleen of a viral lysate-immunized mouse. Antibodies were selected by panning or by enrichment with biotinylated antigen, yielding four different families of antibody fragments. The different types of scFvs were characterized by affinity measurements, by antigen recognition on Western blots, and by pepscan analysis. The epitope of one of the scFvs is located near the residues involved in CypA binding, thereby making it an attractive candidate for therapeutic applications. Comparison of the V gene sequence of this scFV with that of a previously described monoclonal antibody reactive against this immunodominant epitope revealed the usage of the identical combination of VH and Vkappa regions. Thus, this is one of the rare examples in which the original combination in a library-derived antibody fragment was retrieved. After appropriate affinity and format improvements, the best of our recombinant scFvs may form the basis for a sensitive p24 assay as a measure of viral load. In addition, anti-p24 scFvs could be expressed as intracellular antibodies (intrabodies) to aid in the treatment of HIV infections.

* Corresponding author. Mailing address: Functional Biomolecules, Unilever Research Laboratorium Vlaardingen, P.O. Box 114, 3130 AC Vlaardingen, The Netherlands. Phone: (31)-10-4606175. Fax: (31)-10-4605383. E-mail: HANS-DE.HAARD{at}UNILEVER.COM.

Clinical and Diagnostic Laboratory Immunology, September 1998, p. 636-644, Vol. 5, No. 5
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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  • de Haard, H. J., van Neer, N., Reurs, A., Hufton, S. E., Roovers, R. C., Henderikx, P., de Bruine, A. P., Arends, J.-W., Hoogenboom, H. R. (1999). A Large Non-immunized Human Fab Fragment Phage Library That Permits Rapid Isolation and Kinetic Analysis of High Affinity Antibodies. J. Biol. Chem. 274: 18218-18230 [Abstract] [Full Text]  


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