Production of monoclonal antibody against alkaline phosphatase isolated from mouse lymphoid cell line SJL 46, 1985

Shidali, Nathaniel Ndamadu

1980-1989

Alkaline phosphatase (APase) is a cell surface enzyme found to be associated with the membrane of lymphoblastic cells and some lymphoid cell lines. This enzyme appears and disappears in the early differentiation processes of the cells of the immune system (B and T cells). The appearance of APase in pre-T cells seems to preceed that of terminal deoxynucleotide transferase (TdT). The specific role of APase in T cell differention path way is not known. It has been hypothesized that APase acts as an early differentiation marker for pre-T cells. The goal of this research is to produce a monoclonal antibody against APase using SJL-46 cells, a Pre-T cell line, as the source of the enzyme. The monoclonal antibody can then be used to- isolate and purify proteins with APase activity and detect cross-reacting material. At the cellular level, the monoclonal antibody can also be used to identify APase+ cells, remove APase+ cells and sort APase+ cells from a heterogenous mixture of cells. Partial purification of the APase from SJL 46 cells by Nonidet P40 solubilization and butanol extraction yielded a 500-800-fold purification. Mouse myeloma cells SP2/OAgl4, were fused with the spleen cells of Spraque-Dawley rats immunized with the partially purified APase. Using enzyme-antigen-immuno-assay method, eight clones producing monoclonal antibodies against SJL 46 APase were isolated. The objective of the investigation was met by isolating 8 clones secreting anti-APase monoclonal antibody.

text

application/pdf

1985-05-01

thesis

Master of Science (MS)

Atlanta University

Biology

Lumb, Judith Rae

Clark Atlanta University

Georgia--Atlanta

http://hdl.handle.net/20.500.12322/cau.td:1985_shidali_nathaniel_ndamadu