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. 1983 Jun;80(11):3255–3259. doi: 10.1073/pnas.80.11.3255

Monoclonal antibodies to renal brush border membrane maltase: age-associated antigenic alterations.

U Reiss, B Sacktor
PMCID: PMC394019  PMID: 6190174

Abstract

Monoclonal antibodies to maltase (alpha-D-glucoside glucohydrolase, EC 3.2.1.20) from young adult and aged rats were prepared by the hybridoma technique. Four cell lines producing antibodies of the IgG1 subclass to maltase were established. Two, designated 1F12E1 and 8B1G6, produced monoclonal antibodies specific for the catalytically active form of the enzyme found predominantly in enzyme preparations from young animals. The other two clones designated 7G10H3 and 2E1C10 produced monoclonal antibodies that reacted exclusively with an enzymatically inactive form of maltase found mostly in enzyme preparations from aged rats. The increased prevalence of an inactive form of the enzyme in the old rat accounts for the decreased maltase-specific activity previously reported in the senescent rat. The active and inactive maltase species were separated by immunoaffinity chromatography by using the monoclonal antibodies as ligands. The separated forms of the enzyme were not distinguished by NaDodSO4/polyacrylamide gel electrophoresis, peptide mapping of the CNBr-cleaved proteins, and the NH2-terminal residues of these peptides. This study demonstrates the presence of an altered, antigenically distinct enzyme in senescent animals. Critical issues on the mechanism of the aging process may be addressed by application of these findings.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Berger S. J., Sacktor B. Isolation and biochemical characterization of brush borders from rabbit kidney. J Cell Biol. 1970 Dec;47(3):637–645. doi: 10.1083/jcb.47.3.637. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry. 1971 Sep;8(9):871–874. doi: 10.1016/0019-2791(71)90454-x. [DOI] [PubMed] [Google Scholar]
  3. Ey P. L., Prowse S. J., Jenkin C. R. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-sepharose. Immunochemistry. 1978 Jul;15(7):429–436. doi: 10.1016/0161-5890(78)90070-6. [DOI] [PubMed] [Google Scholar]
  4. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  5. Marchalonis J. J., Cone R. E., Santer V. Enzymic iodination. A probe for accessible surface proteins of normal and neoplastic lymphocytes. Biochem J. 1971 Oct;124(5):921–927. doi: 10.1042/bj1240921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. O'Bryan D., Lowenstein L. M. Effect of aging on renal membrane-bound enzyme activities. Biochim Biophys Acta. 1974 Feb 26;339(1):1–9. doi: 10.1016/0005-2736(74)90328-9. [DOI] [PubMed] [Google Scholar]
  7. Reiss U., Sacktor B. Alteration of kidney brush border membrane maltase in aging rats. Biochim Biophys Acta. 1982 Jun 24;704(3):422–426. doi: 10.1016/0167-4838(82)90063-2. [DOI] [PubMed] [Google Scholar]
  8. Reiss U., Sacktor B. Kidney brush border membrane maltase: purification and properties. Arch Biochem Biophys. 1981 Jul;209(2):342–348. doi: 10.1016/0003-9861(81)90290-3. [DOI] [PubMed] [Google Scholar]
  9. Sacktor B. Trehalase and the transport of glucose in the mammalian kidney and intestine. Proc Natl Acad Sci U S A. 1968 Jul;60(3):1007–1014. doi: 10.1073/pnas.60.3.1007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Weiner A. M., Platt T., Weber K. Amino-terminal sequence analysis of proteins purified on a nanomole scale by gel electrophoresis. J Biol Chem. 1972 May 25;247(10):3242–3251. [PubMed] [Google Scholar]
  11. Yuan P. M., Dewan R. N., Zaun M., Thompson R. E., Gracy R. W. Isolation and characterization of triosephosphate isomerase isozymes from human placenta. Arch Biochem Biophys. 1979 Nov;198(1):42–52. doi: 10.1016/0003-9861(79)90393-x. [DOI] [PubMed] [Google Scholar]
  12. de StGroth S. F., Scheidegger D. Production of monoclonal antibodies: strategy and tactics. J Immunol Methods. 1980;35(1-2):1–21. doi: 10.1016/0022-1759(80)90146-5. [DOI] [PubMed] [Google Scholar]

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