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Human Reproduction, Vol. 16, No. 9, 1854-1860, September 2001
© 2001 European Society of Human Reproduction and Embryology

RASA, a recombinant single-chain variable fragment (scFv) antibody directed against the human sperm surface: implications for novel contraceptives

E.J. Norton, A.B. Diekman, V.A. Westbrook, C.J. Flickinger and J.C. Herr,1,*

Department of Cell Biology, Center for Recombinant Gamete Contraceptive Vaccinogens, University of Virginia Health System, Charlottesville, Virginia 22908, USA

BACKGROUND: A recombinant single-chain variable fragment (scFv) antibody was engineered to a tissue-specific carbohydrate epitope located on human sperm agglutination antigen-1 (SAGA-1), a sperm glycoform of CD52. METHODS AND RESULTS: cDNAs encoding the variable regions of the S19 [IgG1{kappa}] monoclonal antibody (mAb) were identified, linked, and cloned into the pCANTAB 5E vector. The recombinant anti-sperm antibody (RASA) was expressed in E. coli HB2151 cells as a 29 kDa monomer and, remarkably, also formed multimers of ~60 and 90 kDa. RASA reacted with the endogenous SAGA-1 antigen by Western blot analysis, labelled the entire human sperm surface by indirect immunofluorescence, and aggregated human spermatozoa in a tangled (head-to-head, head-to-tail, tail-to-tail) pattern of agglutination, as was also observed with the native S19 mAb. CONCLUSIONS: These results demonstrate that active recombinant antibodies can be produced to a tissue-specific carbohydrate epitope on the human sperm surface, thereby opening opportunities for novel contraceptive agents.

Key words: agglutination/antibody engineering/contraception/recombinant antibody/single-chain Fv

1 To whom correspondence should be addressed. E-mail: jch7k{at}virginia.edu

* Dr Herr serves as an officer and member of the Board of Directors of Contra Vac, Inc.


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