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. 1988 Sep;65(1):43–46.

Antigenicity of the carboxyl terminus of insulin: isolation of human insulin-specific monoclonal antibodies.

I H Mirza 1, T J Wilkin 1
PMCID: PMC1385017  PMID: 3053426

Abstract

Monoclonal technology was used to isolate antibodies binding the B30 (carboxy) terminal residue in the polyclonal-provoked immune response to human insulin. Although both spleen and lymph node cell fusions were carried out, only the latter were successful in isolating monoclonal antibodies that bound the carboxy terminal of human insulin. The binding of such antibodies was abolished or diminished by substitutions of the B30 residue. Studies with insulin species variants showed that the molecular binding between antibody and insulin may be critically determined by a subresidue feature, e.g. presence or absence of a single methyl group, as shown by the binding of the monoclonal antibody D10 to human insulin (threonine at B30) but not to rabbit insulin (serine at B30). Such studies are of interest in the study of the molecular basis of antibody-antigen interaction.

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Selected References

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

  1. Axén R., Porath J., Ernback S. Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides. Nature. 1967 Jun 24;214(5095):1302–1304. doi: 10.1038/2141302a0. [DOI] [PubMed] [Google Scholar]
  2. Barcinski M. A., Rosenthal A. S. Immune response gene control of determinant selection. I. Intramolecular mapping of the immunogenic sites on insulin recognized by guinea pig T and B cells. J Exp Med. 1977 Mar 1;145(3):726–742. doi: 10.1084/jem.145.3.726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benjamin D. C., Berzofsky J. A., East I. J., Gurd F. R., Hannum C., Leach S. J., Margoliash E., Michael J. G., Miller A., Prager E. M. The antigenic structure of proteins: a reappraisal. Annu Rev Immunol. 1984;2:67–101. doi: 10.1146/annurev.iy.02.040184.000435. [DOI] [PubMed] [Google Scholar]
  4. Berzofsky J. A. Intrinsic and extrinsic factors in protein antigenic structure. Science. 1985 Sep 6;229(4717):932–940. doi: 10.1126/science.2410982. [DOI] [PubMed] [Google Scholar]
  5. Blundell T. L., Cutfield J. F., Cutfield S. M., Dodson E. J., Dodson G. G., Hodgkin D. C., Mercola D. A. Three-dimensional atomic structure of insulin and its relationship to activity. Diabetes. 1972;21(2 Suppl):492–505. doi: 10.2337/diab.21.2.s492. [DOI] [PubMed] [Google Scholar]
  6. Galfrè G., Milstein C. Preparation of monoclonal antibodies: strategies and procedures. Methods Enzymol. 1981;73(Pt B):3–46. doi: 10.1016/0076-6879(81)73054-4. [DOI] [PubMed] [Google Scholar]
  7. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  8. Marks A., Yip C., Wilson S. Characterization of two epitopes on insulin using monoclonal antibodies. Mol Immunol. 1985 Mar;22(3):285–290. doi: 10.1016/0161-5890(85)90163-4. [DOI] [PubMed] [Google Scholar]
  9. Rathjen D. A., Underwood P. A. Identification of antigenic determinants on insulin recognized by monoclonal antibodies. Mol Immunol. 1986 Apr;23(4):441–450. doi: 10.1016/0161-5890(86)90142-2. [DOI] [PubMed] [Google Scholar]
  10. Schroer J. A., Bender T., Feldmann R. J., Kim K. J. Mapping epitopes on the insulin molecule using monoclonal antibodies. Eur J Immunol. 1983 Sep;13(9):693–700. doi: 10.1002/eji.1830130902. [DOI] [PubMed] [Google Scholar]
  11. Storch M. J., Petersen K. G., Licht T., Kerp L. Recognition of human insulin and proinsulin by monoclonal antibodies. Diabetes. 1985 Aug;34(8):808–811. doi: 10.2337/diab.34.8.808. [DOI] [PubMed] [Google Scholar]
  12. Tainer J. A., Getzoff E. D., Paterson Y., Olson A. J., Lerner R. A. The atomic mobility component of protein antigenicity. Annu Rev Immunol. 1985;3:501–535. doi: 10.1146/annurev.iy.03.040185.002441. [DOI] [PubMed] [Google Scholar]
  13. Thornton J. M., Sibanda B. L. Amino and carboxy-terminal regions in globular proteins. J Mol Biol. 1983 Jun 25;167(2):443–460. doi: 10.1016/s0022-2836(83)80344-1. [DOI] [PubMed] [Google Scholar]
  14. Wilkin T. J., Nicholson S. Autoantibodies against human insulin. Br Med J (Clin Res Ed) 1984 Feb 4;288(6414):349–352. doi: 10.1136/bmj.288.6414.349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Wilkin T., Nicholson S., Casey C. A micro enzyme-linked immunosorbent assay for insulin antibodies in serum. J Immunol Methods. 1985 Jan 21;76(1):185–194. doi: 10.1016/0022-1759(85)90490-9. [DOI] [PubMed] [Google Scholar]

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