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. 1991 Aug;73(4):383–387.

Murine monoclonal anti-Ba antibody that enhances haemolytic activity of factor B.

E Tanaka 1, K Hong 1, T Kinoshita 1, Y Takata 1, H Kozono 1, J Takeda 1, A Yoden 1, K Inoue 1
PMCID: PMC1384564  PMID: 1717366

Abstract

A murine monoclonal antibody (mAb 20-ET) (IgG1, kappa) was selected from a panel of stable hybridomas produced by fusion of P3-X-63-Ag8-U1 (P3UI) myeloma cells with spleen cells from a BALB/c mouse immunized with human factor B. This antibody was shown by the immune blotting method to be directed against the Ba domain of factor B. The haemolytic activity of factor B was enhanced dose-dependently by mAb 20-ET when it was incubated with factor B and EAC4b, 3b cells (sensitized erythrocytes bearing complement fragments C4b and C3b). However, when the antibody was added after factor B had been bound to EAC4b,3b cells and the cells had been washed, it caused little enhancement of the haemolytic activity. The enhancing effect of this antibody was not due to its stabilization of the C3b-B complex, because EAC4b,3b dissociated from mAb 20-ET-bound factor B complexes rather more readily than from uncomplexed factor B. The presence of mAb 20-ET in the reaction mixture caused and maintained a much higher steady-state level of binding of factor B with EAC4b,3b cells than that in its absence. Factor P caused delayed dissociation of mAb-bound factor B from EAC4b,3b cells, thus enhancing the haemolytic activity of factor B bound with the mAb.

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Selected References

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

  1. Bolotin C., Morris S., Tack B., Prahl J. Purification and structural analysis of the fourth component of human complement. Biochemistry. 1977 May 3;16(9):2008–2015. doi: 10.1021/bi00628a039. [DOI] [PubMed] [Google Scholar]
  2. Daha M. R., Deelder A. M., Van Es L. A. Stabilization of the amplification convertase of complement by monoclonal antibodies directed against human factor B. J Immunol. 1984 May;132(5):2538–2542. [PubMed] [Google Scholar]
  3. Daha M. R., Fearon D. T., Austen K. F. Isolation of alternative pathway C3 convertase containing uncleaved B and formed in the presence of C3 nephritic factor (C3neF). J Immunol. 1976 Feb;116(2):568–570. [PubMed] [Google Scholar]
  4. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. J Immunol. 1972 Jul;109(1):129–135. [PubMed] [Google Scholar]
  5. Hammer C. H., Wirtz G. H., Renfer L., Gresham H. D., Tack B. F. Large scale isolation of functionally active components of the human complement system. J Biol Chem. 1981 Apr 25;256(8):3995–4006. [PubMed] [Google Scholar]
  6. Hong K., Kinoshita T., Dohi Y., Inoue K. Effect of trypsinization on the activity of human factor H. J Immunol. 1982 Aug;129(2):647–652. [PubMed] [Google Scholar]
  7. Hong K., Kinoshita T., Miyazaki W., Izawa T., Inoue K. An anticomplementary agent, K-76 monocarboxylic acid: its site and mechanism of inhibition of the complement activation cascade. J Immunol. 1979 Jun;122(6):2418–2423. [PubMed] [Google Scholar]
  8. Inoue K., Nelson R. A., Jr The isolation and characterization of a new component of hemolytic complement, C'3e. J Immunol. 1965 Aug;95(2):355–367. [PubMed] [Google Scholar]
  9. Kerr M. A., Porter R. R. The purification and properties of the second component of human complement. Biochem J. 1978 Apr 1;171(1):99–107. doi: 10.1042/bj1710099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Lambris J. D., Müller-Eberhard H. J. Isolation and characterization of a 33,000-dalton fragment of complement Factor B with catalytic and C3b binding activity. J Biol Chem. 1984 Oct 25;259(20):12685–12690. [PubMed] [Google Scholar]
  12. Lesavre P. H., Hugli T. E., Esser A. F., Müller-Eberhard H. J. The alternative pathway C3/C5 convertase: chemical basis of factor B activation. J Immunol. 1979 Aug;123(2):529–534. [PubMed] [Google Scholar]
  13. Markwell M. A., Fox C. F. Surface-specific iodination of membrane proteins of viruses and eucaryotic cells using 1,3,4,6-tetrachloro-3alpha,6alpha-diphenylglycoluril. Biochemistry. 1978 Oct 31;17(22):4807–4817. doi: 10.1021/bi00615a031. [DOI] [PubMed] [Google Scholar]
  14. Medicus R. G., Esser A. F., Fernandez H. N., Müller-Eberhard H. J. Native and activated properdin: interconvertibility and identity of amino- and carboxy-terminal sequences. J Immunol. 1980 Feb;124(2):602–606. [PubMed] [Google Scholar]
  15. Pryzdial E. L., Isenman D. E. Alternative complement pathway activation fragment Ba binds to C3b. Evidence that formation of the factor B-C3b complex involves two discrete points of contact. J Biol Chem. 1987 Feb 5;262(4):1519–1525. [PubMed] [Google Scholar]
  16. Tack B. D., Prahl J. W. Third component of human complement: purification from plasma and physicochemical characterization. Biochemistry. 1976 Oct 5;15(20):4513–4521. doi: 10.1021/bi00665a028. [DOI] [PubMed] [Google Scholar]
  17. Tamura N., Nelson R. A., Jr The purification and reactivity of the first component of complement from guinea pig, human and canine sera. J Immunol. 1968 Dec;101(6):1333–1345. [PubMed] [Google Scholar]
  18. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ueda A., Kearney J. F., Roux K. H., Volanakis J. E. Probing functional sites on complement protein B with monoclonal antibodies. Evidence for C3b-binding sites on Ba. J Immunol. 1987 Feb 15;138(4):1143–1149. [PubMed] [Google Scholar]
  20. Vogt W., Dames W., Schmidt G., Dieminger L. Complement activation by the properdin system: formation of a stoichiometric. C3 cleaving complex of properdin factor B with C36. Immunochemistry. 1977 Mar;14(3):201–205. doi: 10.1016/0019-2791(77)90195-1. [DOI] [PubMed] [Google Scholar]

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