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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1985 Jun;82(11):3839–3843. doi: 10.1073/pnas.82.11.3839

Polymorphism of normal factor IX detected by mouse monoclonal antibodies.

A Wallmark, R Ljung, I M Nilsson, L Holmberg, U Hedner, M Lindvall, H O Sjögren
PMCID: PMC397883  PMID: 3873655

Abstract

Hemophilia B is an X-chromosomal recessive disease due to deficiency of coagulation factor IX. Three monoclonal antibodies against factor IX were prepared and used to develop immunoradiometric assays (IRMAs) of factor IX antigen (IX-Ag). IX-Ag was measured in 65 normal individuals with one IRMA based on polyclonal anti-IX antibodies and two IRMAs based on three monoclonal anti-IX antibodies. One of the monoclonal antibodies differed in specificity since it neutralized less than 50% of the clotting activity of factor IX (IX-C), whereas the other two monoclonal antibodies neutralized 80-95%. When the former antibody was used as the solid phase in IRMA, two groups of normal individuals were distinguished: group A with measurable IX-Ag, and group B without demonstrable IX-Ag. There were no differences between the groups either in IX-C or in IX-Ag measured with polyclonal antibodies. A subgroup comprising only women could be distinguished in group A, in whom intermediate IX-Ag concentrations were found. Family studies showed the group B variant of normal factor IX to be transmitted according to the pattern of X-linked recessive inheritance. The allelic frequency of group A was 0.66, and that of group B was 0.34.

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Selected References

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

  1. Amphlett G. W., Kisiel W., Castellino F. J. The interaction of Ca2+ with human Factor IX. Arch Biochem Biophys. 1981 May;208(2):576–585. doi: 10.1016/0003-9861(81)90546-4. [DOI] [PubMed] [Google Scholar]
  2. Anson D. S., Choo K. H., Rees D. J., Giannelli F., Gould K., Huddleston J. A., Brownlee G. G. The gene structure of human anti-haemophilic factor IX. EMBO J. 1984 May;3(5):1053–1060. doi: 10.1002/j.1460-2075.1984.tb01926.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BRINKHOUS K. M., LANGDELL R. D., PENICK G. D., GRAHAM J. B., WAGNER R. H. Newer approaches to the study of hemophilia and hemophilioid states. J Am Med Assoc. 1954 Feb 6;154(6):481–486. doi: 10.1001/jama.1954.02940400019005. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Buchanan D., Kamarck M., Ruddle N. H. Development of a protein A enzyme immunoassay for use in screening hybridomas. J Immunol Methods. 1981;42(2):179–185. doi: 10.1016/0022-1759(81)90147-2. [DOI] [PubMed] [Google Scholar]
  6. Camerino G., Grzeschik K. H., Jaye M., De La Salle H., Tolstoshev P., Lecocq J. P., Heilig R., Mandel J. L. Regional localization on the human X chromosome and polymorphism of the coagulation factor IX gene (hemophilia B locus). Proc Natl Acad Sci U S A. 1984 Jan;81(2):498–502. doi: 10.1073/pnas.81.2.498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Denson K. W., Biggs R., Mannucci P. M. An investigation of three patients with Christmas disease due to an abnormal type of factor IX. J Clin Pathol. 1968 Mar;21(2):160–165. doi: 10.1136/jcp.21.2.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Di Scipio R. G., Hermodson M. A., Yates S. G., Davie E. W. A comparison of human prothrombin, factor IX (Christmas factor), factor X (Stuart factor), and protein S. Biochemistry. 1977 Feb 22;16(4):698–706. doi: 10.1021/bi00623a022. [DOI] [PubMed] [Google Scholar]
  9. Di Scipio R. G., Kurachi K., Davie E. W. Activation of human factor IX (Christmas factor). J Clin Invest. 1978 Jun;61(6):1528–1538. doi: 10.1172/JCI109073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fagerhol M. K., Laurell C. B. The Pi system-inherited variants of serum alpha 1-antitrypsin. Prog Med Genet. 1970;7:96–111. [PubMed] [Google Scholar]
  11. Fujikawa K., Legaz M. E., Kato H., Davie E. W. The mechanism of activation of bovine factor IX (Christmas factor) by bovine factor XIa (activated plasma thromboplastin antecedent). Biochemistry. 1974 Oct 22;13(22):4508–4516. doi: 10.1021/bi00719a006. [DOI] [PubMed] [Google Scholar]
  12. GRUBB R. Agglutination of erythrocytes coated with incomplete anti-Rh by certain rheumatoid arthritic sera and some other sera; the existence of human serum groups. Acta Pathol Microbiol Scand. 1956;39(3):195–197. [PubMed] [Google Scholar]
  13. Giannelli F., Anson D. S., Choo K. H., Rees D. J., Winship P. R., Ferrari N., Rizza C. R., Brownlee G. G. Characterisation and use of an intragenic polymorphic marker for detection of carriers of haemophilia B (factor IX deficiency). Lancet. 1984 Feb 4;1(8371):239–241. doi: 10.1016/s0140-6736(84)90122-3. [DOI] [PubMed] [Google Scholar]
  14. Giannelli F., Choo K. H., Rees D. J., Boyd Y., Rizza C. R., Brownlee G. G. Gene deletions in patients with haemophilia B and anti-factor IX antibodies. Nature. 1983 May 12;303(5913):181–182. doi: 10.1038/303181a0. [DOI] [PubMed] [Google Scholar]
  15. Goding J. W. Antibody production by hybridomas. J Immunol Methods. 1980;39(4):285–308. doi: 10.1016/0022-1759(80)90230-6. [DOI] [PubMed] [Google Scholar]
  16. Goodall A. H., Kemble G., O'Brien D. P., Rawlings E., Rotblat F., Russell G. C., Janossy G., Tuddenham E. G. Preparation of factor IX deficient human plasma by immunoaffinity chromatography using a monoclonal antibody. Blood. 1982 Mar;59(3):664–670. [PubMed] [Google Scholar]
  17. Holmberg L., Gustavii B., Cordesius E., Kristoffersson A. C., Ljung R., Löfberg L., Strömberg P., Nilsson I. M. Prenatal diagnosis of hemophilia B by an immunoradiometric assay of factor IX. Blood. 1980 Sep;56(3):397–401. [PubMed] [Google Scholar]
  18. Holmberg L., Nilsson I. M. Immunologic studies in haemophilia A. Scand J Haematol. 1973;10(1):12–16. doi: 10.1111/j.1600-0609.1973.tb00032.x. [DOI] [PubMed] [Google Scholar]
  19. Jaye M., de la Salle H., Schamber F., Balland A., Kohli V., Findeli A., Tolstoshev P., Lecocq J. P. Isolation of a human anti-haemophilic factor IX cDNA clone using a unique 52-base synthetic oligonucleotide probe deduced from the amino acid sequence of bovine factor IX. Nucleic Acids Res. 1983 Apr 25;11(8):2325–2335. doi: 10.1093/nar/11.8.2325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kurachi K., Davie E. W. Isolation and characterization of a cDNA coding for human factor IX. Proc Natl Acad Sci U S A. 1982 Nov;79(21):6461–6464. doi: 10.1073/pnas.79.21.6461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. LYON M. F. Sex chromatin and gene action in the mammalian X-chromosome. Am J Hum Genet. 1962 Jun;14:135–148. [PMC free article] [PubMed] [Google Scholar]
  22. Lamme S., Wallmark A., Holmberg L., Nilsson I. M., Sjögren H. O. The use of monoclonal antibodies in measuring factor VIII/von Willebrand factor. Scand J Clin Lab Invest. 1985 Feb;45(1):17–26. doi: 10.3109/00365518509160967. [DOI] [PubMed] [Google Scholar]
  23. Laurell C. B. Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal Biochem. 1966 Apr;15(1):45–52. doi: 10.1016/0003-2697(66)90246-6. [DOI] [PubMed] [Google Scholar]
  24. Ljung R., Holmberg L. Genetic variants of haemophilia B detected by immunoradiometric assay: implications for prenatal diagnosis. Pediatr Res. 1982 Mar;16(3):256–258. doi: 10.1203/00006450-198203000-00019. [DOI] [PubMed] [Google Scholar]
  25. NENNSTIEL H. J., BECHT T. Uber das erbliche Auftreten einer Albuminspaltung im Elektrophoresedlagramm. Klin Wochenschr. 1957 Jul 1;35(13):689–689. doi: 10.1007/BF01481274. [DOI] [PubMed] [Google Scholar]
  26. NILSSON I. M., BLOMBACK M., RAMGREN O. Haemophilia in Sweden. I. Coagulation studies. Acta Med Scand. 1961 Dec;170:665–682. [PubMed] [Google Scholar]
  27. OUDIN J. Réaction de précipitation spécifique entre des sérums d'animaux de même espèce. C R Hebd Seances Acad Sci. 1956 May 14;242(20):2489–2490. [PubMed] [Google Scholar]
  28. Parekh V. R., Mannucci P. M., Ruggeri Z. M. Immunological heterogeneity of haemophilia B: a multicentre study of 98 kindreds. Br J Haematol. 1978 Dec;40(4):643–655. doi: 10.1111/j.1365-2141.1978.tb05840.x. [DOI] [PubMed] [Google Scholar]
  29. Peake I. R., Furlong B. L., Bloom A. L. Carrier detection by direct gene analysis in a family with haemophilia B (factor IX deficiency). Lancet. 1984 Feb 4;1(8371):242–243. doi: 10.1016/s0140-6736(84)90123-5. [DOI] [PubMed] [Google Scholar]
  30. Roberts H. R., Grizzle J. E., McLester W. D., Penick G. D. Genetic variants of hemophilia B: detection by means of a specific PTC inhibitor. J Clin Invest. 1968 Feb;47(2):360–365. doi: 10.1172/JCI105732. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. SMITHIES O. Variations in human serum beta-globulins. Nature. 1957 Dec 28;180(4600):1482–1483. doi: 10.1038/1801482a0. [DOI] [PubMed] [Google Scholar]
  32. SMITHIES O. Zone electrophoresis in starch gels: group variations in the serum proteins of normal human adults. Biochem J. 1955 Dec;61(4):629–641. doi: 10.1042/bj0610629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Smith K. J., Ono K. Monoclonal antibodies to factor IX: characterization and use in immunoassays for factor IX. Thromb Res. 1984 Jan 15;33(2):211–224. doi: 10.1016/0049-3848(84)90182-8. [DOI] [PubMed] [Google Scholar]
  34. Theodorsson B., Hedner U., Nilsson I. M., Kisiel W. A technique for specific removal of factor IX alloantibodies from human plasma: partial characterization of the alloantibodies. Blood. 1983 May;61(5):973–981. [PubMed] [Google Scholar]
  35. Thompson A. R. Monoclonal antibody to an epitope on the heavy chain of factor IX missing in three hemophilia-B patients. Blood. 1983 Nov;62(5):1027–1034. [PubMed] [Google Scholar]
  36. Thorell J. I., Johansson B. G. Enzymatic iodination of polypeptides with 125I to high specific activity. Biochim Biophys Acta. 1971 Dec 28;251(3):363–369. doi: 10.1016/0005-2795(71)90123-1. [DOI] [PubMed] [Google Scholar]
  37. Wallmark A., Alm R., Eriksson S. Monoclonal antibody specific for the mutant PiZ alpha 1-antitrypsin and its application in an ELISA procedure for identification of PiZ gene carriers. Proc Natl Acad Sci U S A. 1984 Sep;81(18):5690–5693. doi: 10.1073/pnas.81.18.5690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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