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Developing methods for antibody-mediated enrichment and capture of spermatozoa from complex cell mixtures for forensics applications

Jackson, Monique Bailey (2019) Developing methods for antibody-mediated enrichment and capture of spermatozoa from complex cell mixtures for forensics applications. Honours thesis, Murdoch University.

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Cases of sexual assault have steadily increased over the past years. Sexual assault samples are complex with vaginal epithelial cells as the main contributor to the sample, while spermatozoa are vast in number. There a variety of techniques such as differential extraction and laser capture microdissection that exist to isolate and capture spermatozoa for DNA profiling. These methods are tedious and are not always efficient. Anti-sperm antibodies can be used to target spermatozoa by binding to antigens localised on the surface of the acrosome. This study tested the binding capacity of a variety of anti-sperm antibodies to fresh and aged spermatozoa. The binding ability of the α-SPAM1 primary antibody to spermatozoa eluted from Copan rayon swabs was assessed to evaluate that the number of spermatozoa stained was sufficient for STR analysis. This study showed that α-SPAM1 and α-SPACA1 stained more than >30% of cells in fresh and aged spermatozoa. This study found that 39% of aged spermatozoa were stained with a combination of α-SPAM1 and α-SPACA1. It was found that Copan rayon swabs were efficient at absorbing spermatozoa but only released <50% of cells. Copan rayon swabs retain 33% of initial spermatozoa consumed. The study found that despite the high retention capacity of Copan rayon swabs, sufficient numbers of spermatozoa were eluted and stained with α-SPAM1 at levels compatible with DNA techniques such as STR analysis. In conclusion, this study shows that in future studies, α-SPAM1 and α-SPACA1 could be used to in conjunction with immunomagnetic beads to extract spermatozoa from sexual assault samples.

Item Type: Thesis (Honours)
Murdoch Affiliation(s): School of Veterinary and Life Sciences
United Nations SDGs: Goal 9: Industry, Innovation and Infrastructure
Goal 16: Peace, Justice and Strong Institutions
Supervisor(s): Chapman, Brendan and Currie, Andrew
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