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. 1979 Jul;64(1):337–341. doi: 10.1172/JCI109459

Production of hybridomas secreting monoclonal antibodies against the lympholine osteoclast activating factor.

R A Luben, M A Mohler, G E Nedwin
PMCID: PMC372124  PMID: 221547

Abstract

The human lympholine osteoclast activating factor (OAF) is thought to be involved in several bone-destroying diseases. The current studies were designed to produce monoclonal antibodies against OAF for use in the subsequent design of immunoassays for OAF in clinical samples. Spleen cells from mice immunized with purified human OAF were hybridized with mouse plasmacytoma cells in vitro to yield hybridomas. Several clones of these hybridomas secreted into the culture medium antibodies, which neutralized the biological activity of OAF at dilutions as high as 1:100,000 relative to the initial culture medium. These antibodies did not interfere with the activities of parathyroid hormone in the same systems. These results represent the first report of monoclonal antibodies against a human lympholine, and validate the concept that hybridoma production is a useful technique for developing antibodies against weak or scarce antigens.

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Selected References

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

  1. Galfre G., Howe S. C., Milstein C., Butcher G. W., Howard J. C. Antibodies to major histocompatibility antigens produced by hybrid cell lines. Nature. 1977 Apr 7;266(5602):550–552. doi: 10.1038/266550a0. [DOI] [PubMed] [Google Scholar]
  2. Goding J. W. Use of staphylococcal protein A as an immunological reagent. J Immunol Methods. 1978;20:241–253. doi: 10.1016/0022-1759(78)90259-4. [DOI] [PubMed] [Google Scholar]
  3. Horton J. E., Raisz L. G., Simmons H. A., Oppenheim J. J., Mergenhagen S. E. Bone resorbing activity in supernatant fluid from cultured human peripheral blood leukocytes. Science. 1972 Sep 1;177(4051):793–795. doi: 10.1126/science.177.4051.793. [DOI] [PubMed] [Google Scholar]
  4. Klinger H. P. Rapid processing of primary embryonic tissues for chromosome banding pattern analysis. Cytogenetics. 1972;11(5):424–435. doi: 10.1159/000130208. [DOI] [PubMed] [Google Scholar]
  5. Littlefield J. W. The use of drug-resistant markers to study the hybridization of mouse fibroblasts. Exp Cell Res. 1966 Jan;41(1):190–196. doi: 10.1016/0014-4827(66)90558-1. [DOI] [PubMed] [Google Scholar]
  6. Luben R. A., Cohn D. V. Effects of parathormone and calcitonin on citrate and hyaluronate metabolism in cultured bone. Endocrinology. 1976 Feb;98(2):413–419. doi: 10.1210/endo-98-2-413. [DOI] [PubMed] [Google Scholar]
  7. Luben R. A., Mundy G. R., Trummel C. L., Raisz L. G. Partial purification of osteoclast-activating factor from phytohemagglutinin-stimulated human leukocytes. J Clin Invest. 1974 May;53(5):1473–1480. doi: 10.1172/JCI107696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Luben R. A. Purification of a lymphokine: osteoclast activating factor from human tonsil lymphocytes. Biochem Biophys Res Commun. 1978 Sep 14;84(1):15–22. doi: 10.1016/0006-291x(78)90256-5. [DOI] [PubMed] [Google Scholar]
  9. Margulies D. H., Kuehl W. M., Scharff M. D. Somatic cell hybridization of mouse myeloma cells. Cell. 1976 Jul;8(3):405–415. doi: 10.1016/0092-8674(76)90153-7. [DOI] [PubMed] [Google Scholar]
  10. Mundy G. R., Luben R. A., Raisz L. G., Oppenheim J. J., Buell D. N. Bone-resorbing activity in supernatants from lymphoid cell lines. N Engl J Med. 1974 Apr 18;290(16):867–871. doi: 10.1056/NEJM197404182901601. [DOI] [PubMed] [Google Scholar]
  11. Mundy G. R., Raisz L. G., Cooper R. A., Schechter G. P., Salmon S. E. Evidence for the secretion of an osteoclast stimulating factor in myeloma. N Engl J Med. 1974 Nov 14;291(20):1041–1046. doi: 10.1056/NEJM197411142912001. [DOI] [PubMed] [Google Scholar]
  12. Stern P. L., Willison K. R., Lennox E., Galfrè G., Milstein C., Secher D., Ziegler A. Monoclonal antibodies as probes for differentiation and tumor-associated antigens: a Forssman specificity on teratocarcinoma stem cells. Cell. 1978 Aug;14(4):775–783. doi: 10.1016/0092-8674(78)90333-1. [DOI] [PubMed] [Google Scholar]
  13. Williams A. F., Galfrè G., Milstein C. Analysis of cell surfaces by xenogeneic myeloma-hybrid antibodies: differentiation antigens of rat lymphocytes. Cell. 1977 Nov;12(3):663–673. doi: 10.1016/0092-8674(77)90266-5. [DOI] [PubMed] [Google Scholar]

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