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Development of a Novel Method for the Identification of Human B Cells Producing Antibodies against Staphylococcus aureus Type 8 Capsule

Gaurav, Rahul
http://rave.ohiolink.edu/etdc/view?acc_num=ysu1409846265

Abstract Details

2014, Master of Science in Biological Sciences, Youngstown State University, Department of Biological Sciences and Chemistry.
Staphylococcus aureus infections are the primary cause of nosocomial infections worldwide. Increasing antibiotic resistance in S. aureus requires the development of alternative treatment methods. It has been shown that mice injected with homologous antibodies against Staphylococcus aureus had much lower fatality, when compared to control mice infected with S. aureus. Although mouse monoclonal antibodies against S. aureus have been developed, injection into human subjects results in the production of human anti-murine antibodies in the host. The long term goal of this project is the production of human antibodies against S. aureus. Towards this goal, S. aureus type 8 (Wright strain) was grown in Columbia broth containing 2% sodium chloride, harvested and autoclave. It was then treated with nucleases, pronase and sodium periodate and the carbohydrate purified using DEAE ion exchange chromatography and Sephadex S-300 size-exclusion chromatography. The purified sample contained reducing carbohydrate that was reactive with capsule specific antibodies, but no detectable protein or teichoic acid. Biotin hydrazide was used to biotinylate the carbohydrate sample and binding of anti-capsular antibodies to the biotinylated product was tested using an ELISA. We found that the use of PBS + 0.05% Tween-20 + 0.1% BSA as the sample buffer caused false positive results, which was rectified by the use of casein based sample buffer. In these studies, we have developed an ELISA which can be used to test antibody binding to biotinylated capsular carbohydrate.
Diana Fagan, PhD (Advisor)
David Asch, PhD (Committee Member)
Jonathan Caguiat, PhD (Committee Member)
101 p.
Biology; Cellular Biology; Immunology; Microbiology
Staphylococcus aureus; Capsule; Type 8; ELISA

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Citations

  • Gaurav, R. (2014). Development of a Novel Method for the Identification of Human B Cells Producing Antibodies against Staphylococcus aureus Type 8 Capsule [Master's thesis, Youngstown State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1409846265

    APA Style (7th edition)

  • Gaurav, Rahul. Development of a Novel Method for the Identification of Human B Cells Producing Antibodies against Staphylococcus aureus Type 8 Capsule. 2014. Youngstown State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ysu1409846265.

    MLA Style (8th edition)

  • Gaurav, Rahul. "Development of a Novel Method for the Identification of Human B Cells Producing Antibodies against Staphylococcus aureus Type 8 Capsule." Master's thesis, Youngstown State University, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1409846265

    Chicago Manual of Style (17th edition)

ysu1409846265
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© 2014, all rights reserved.
This open access ETD is published by Youngstown State University and OhioLINK.