This awardee has received supplemental funding. This award detail page includes information about both the original award and supplemental awards.
Description of original award (Fiscal Year 2004, $305,532)
Project summary for 2000-IJ-CX-K013
THe objective of this research project is to develop a sperm cell selection device suitable for forensic DNA analysis of the "male component" in sexual assault evidence. The device and its companion mehthodologies will be compatible with high throughput automation employing PCR of short tandem repeat loci. The device will provide an enabling technology to assist the development of the Combined DNA Index System (CODIS).
Monoclonal antibodies and recombinant mini-antibodies will produced against a unique human sperm surface carbohydrates epitope,sperm agglutination antigen-1(SAGA-1).This antigen is synthesized in the principal cells of the epididymis, is specific to the male reproductive trac of humans and higher primates and is inserted by way of a GPI anchor into all domains of the human sperm surface: Head,midpiece and tail. This antigen thus offers an excellent target for developing methods for separating sperm
cells from other biological materials.
Immuno-reagents to SAGA-1 will be coupled to magnetic beads and magnetite particles.Methodologies employing these prototype devices, including single sperm PCR, will be developed and standardized to yield differential separation of male germ cell DNA from DNA of other cell types.
The microscopic identification of sperm in sexual assault evidence can be labor intensive and time consuming. During the initial phase of this project, fluorescently labeled anti-sperm antibodies were developed. These antibodies (called "Sperm Paints") have the capability to bind to sperm and make them easily visible using a fluorescent microscope. The ability of the Sperm Paints to bind to and illuminate sperm heads and tails in simulated sexual assault evidence that has been stored for prolonged periods has also been demonstrated. The goal of this supplemental funding is to complete the development and optimization of a protocol for using the Sperm Paints for microscopic sperm searches, and to perform testing and validation of the Sperm Paints through collaborative studies with crime labs.
1. Optimize sperm binding by a cocktail of fluorescent monoclonal anti-sperm antibodies (Sperm Paint) which bind to sperm specific proteins located in different domains of the sperm head and tail.
2. Determine the binding characteristics of Sperm Paint to simulated evidence obtained 24, 48, 72 and 96 hours post-coitus.
3. Collaborate with crime labs to evaluate and validate the Sperm Paints.
4. Develop an optimized protocol for Sperm Paint use.