Purification and Characterization of an Anti-Basigin Monoclonal Antibody: Application to the Study of Human Uterine Cells and Tissues

<p>Between 1990 and 2017, the global rate of female infertility increased by over 15% (Sun et al., 2019). An embryo's failure to implant into the uterus is the primary cause of early pregnancy loss. Several studies have identified molecules necessary for successful implantation, but their function during implantation remains poorly understood (S. Zhang et al., 2013). The cell surface glycoprotein basigin-2 is necessary for embryo implantation in the mouse and plays a role in several cellular functions, including cell communication and extracellular matrix remodeling (Chen et al., 2009; K. Li & Nowak, 2020). <em>In vitro</em> studies using human uterine cells demonstrate that basigin-2 regulates the expression and secretion of enzymes called matrix metalloproteinases, which are critical for successful implantation in the mouse (Belton et al., 2008). This thesis aimed to purify and characterize a monoclonal antibody raised against a recombinant form of basigin-2, then measure the expression of basigin-2 proteins in human uterine cells and tissues. The results indicate the P2D7 mAb specifically labeled basigin-2 expression in thin sections of human uterine tissues, and labeling was inhibited by the inclusion of the recombinant basigin protein in a competition assay to show specificity. Immunoblot analysis using the P2D7 mAb demonstrated the specific labeling of human basigin-2 in HESC lysates. The antibody additionally detects basigin in HESCs <em>in vitro</em>. The characterization of the P2D7 mAb supports its use in future studies of the role of basigin-2 in the molecular mechanisms of uterine reproductive function resulting in successful embryo implantation.</p>

Name thesis:degree_name

Master of Science

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Biology

Year dc:date.available

2022

Author dc:creator

• Barna, Jeannine M

Contributors dc:contributor

• Dr. Robert Belton

• reproduction
• immunohistochemistry
• immunocytochemistry
• immunoblot
• endometrium
• Cell and Developmental Biology

Repository record dc:identifier

https://commons.nmu.edu/theses/731

OAI identifier oai:identifier

oai:commons.nmu.edu:theses-1773