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NCJ Number: NCJ 241076     Find in a Library
Title: RASA, A Recombinant Single-Chain Variable Fragment (scFv) Antibody Directed Against the Human Sperm Surface: Implications for Noval Contraceptives
Journal: Human Reproduction  Volume:16  Issue:9  Dated:2001  Pages:1854 to 1860
Author(s): E.J. Norton ; A.B. Diekman ; V.A. Westbrook ; C.J. Flickinger ; J.C. Herr
Date Published: 2001
Page Count: 7
Sponsoring Agency: Andrew W Mellon Foundation
United States of America

National Institute of Justice
US Department of Justice
Office of Justice Programs
United States of America

Fogarty International Ctr
United States of America
Grant Number: 2000-IJ-CX-K013;NIH HD U54 29099;R01 HDD35523;D43 TW/HD00654;R43 HD 35771;BIO-98-002;CIG-97-15
Type: Report (Study/Research)
Language: English
Country: United States of America
Annotation: 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, demonstrating that active recombinant antibodies can be produced to a tissue-specific carbohydrate epitope on the human sperm surface, thereby creating opportunities for novel contraceptive agents.
Abstract: The described materials and methods generated RASA, a recombinant scFv mini-antibody form of the murine anti-sperm mAb S19. Using recombinant DNA techniques, cDNAs encoding the variable regions of the S19 heavy and light chains were isolated from the parental MHS-8 hybridoma, linked to one another, ligated into an expression vector, and expressed in E. coli. Identification of the correct light chain cDNA was initially problematic due to a frame shift in the cDNA generated by the RPAS kit primers and was corrected through the generation of a unique primer. Use of the Van911 restriction enzyme to delete cDNAs encoding the aberrant endogenous light chain was a novel technique that expedited cloning of the authentic scFv. Both techniques may be of use to other researchers generating recombinant antibodies from murine hybridomas with homologous sequences to RASA. Over the past decade, the use of monoclonal antibodies has been expanded to a variety of clinical applications (Hollinger and Hoogenboom, 1998; Hudson, 1998, 1999); however, adverse effects, such as binding interference, caused by using whole murine antibodies in humans, and the expense of tissue culture have presented obstacles to their use. Recombinant antibody technology has presented a solution to both of these impediments, because the generation of recombinant antibody fragments, such as scFvs, not only removes the majority of the immunogenic murine sequence, but may also be more economical. 6 figures and 45 references
Main Term(s): Forensics/Forensic Sciences
Index Term(s): Biological influences ; Abortion ; Adolescent pregnancy ; NIJ grant-related documents
   
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https://www.ncjrs.gov/App/Publications/abstract.aspx?ID=263164

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