A Monoclonal Antibody to Intestinal Alkaline Phosphatase Made Against D98/AH-2 (HeLa) Cells
- 11 December 2008
- journal article
- research article
- Published by Wiley in Tissue Antigens
- Vol. 17 (3), 303-312
- https://doi.org/10.1111/j.1399-0039.1981.tb00705.x
Abstract
A monoclonal antibody (AAP1) to human intestinal alkaline phosphatase (ALP) was produced by immunizing a mouse with D98/AH-2 (HeLa) cells, which produce the enzyme ectopically. The antibody, which did not inhibit enzyme activity using p-nitrophenyl phosphate as the substrate, was of the IgG2A class and did not show complement-dependent cytotoxicity. In trace binding assays, AAP1 bound only to cells that expressed an intestinal-like form of human ALP, including some human intraspecific (D98/AH-2 .times. human lymphocyte or fibroblast) hybrids. Immunoprecipitation of immune complexes from cell-free extracts of D98/AH-2 cells, using protein A containing Staphylococcus aureus and AAP1 antibody, resulted in precipitation of all the ALP activity. The precipitated material had a subunit MW of 80,000 daltons, as estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. In non-denaturing conditions, AAP1 antibody prevented ALP activity migration into the gel when cell-free extracts were made from human adult or fetal intestine, or D98/AH-2 cells. Similarly, AAP1 could be used to precipitate ALP activity from these extracts but not from extracts of human liver, kidney or placenta.Keywords
This publication has 31 references indexed in Scilit:
- Human cell lines expressing intestinal alkaline phosphatase.Proceedings of the National Academy of Sciences, 1979
- Characterization of a monoclonal anti‐β2‐microglobulin antibody and its use in the genetic and biochemical analysis of major histocompatibility antigensEuropean Journal of Immunology, 1979
- Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens-new tools for genetic analysisCell, 1978
- The Genetic Control of HLA‐A and B Antigens in Somatic Cell Hybrids: Requirement for β2 MicroglobulinTissue Antigens, 1978
- Heterogeneity of alkaline phosphatases in different HeLa linesSomatic Cell and Molecular Genetics, 1978
- Structural evidence that human liver and placental alkaline phosphatase isoenzymes are coded by different genes.Proceedings of the National Academy of Sciences, 1976
- Human intraspecific somatic cell hybrids: A genetic and karyotypic analysis of crosses between lymphocytes and D98/AH-2Somatic Cell and Molecular Genetics, 1975
- GENE EXPRESSION IN SOMATIC CELL HYBRIDSAnnual Review of Genetics, 1974
- Perspectives on alkaline phosphatase isoenzymesAmerican Journal Of Medicine, 1974
- Properties of placental alkaline phosphatase. I. Molecular size and electrical charge of the various electrophoretic components of the six common phenotypesBiochemical Genetics, 1970