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Human Genetics

, Volume 70, Issue 2, pp 163–167 | Cite as

Subtyping of haptoglobin — Presentation of a new method

  • Brita Teige
  • Bjørnar Olaisen
  • Lilian Pedersen
Original Investigations

Summary

A method is described for large scale routine phenotyping of haptoglobin (Hp) which allows complete subtyping without prior purification of the Hp molecule. The procedure includes polyacrylamide gel isoelectric focusing of reduced, neuraminidase treated serum or plasma samples, and nitrocellulose blots developed with the immunoperoxidase technique. Different variables including sample treatment, electrofocusing, blotting procedures, and immunoperoxidase visualization are discussed.

Characteristic α-chain patterns allow identification of the common allotypes 2FS, 2SS, 2FF, IS, IF, and Johnson. Isoelectric variations in the β-chain may also be recognized. For comparison, two-dimensional Hp-patterns are presented. The results from concurrent typing of 600 samples by ordinary starch gel electrophoresis and by the described isofocusing technique, are evaluated.

Keywords

Starch Polyacrylamide Metabolic Disease Plasma Sample Sample Treatment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Anderson NG, Anderson NL (1978a) Analytical techniques for cell fractions. XXI. Two-dimensional analysis of serum and tissue proteins: Multiple isoelectric focusing. Anal Biochem 85:331–340CrossRefGoogle Scholar
  2. Anderson NL, Anderson NG (1978b) Analytical techniques for cell fractions. XXII. Two-dimensional analysis of serum and tissue proteins: Multiple gradient-slab gel electrophoresis. Anal Biochem 85:341–354CrossRefGoogle Scholar
  3. Anderson NG, Anderson NL, Tollaksen SL (1979) Operation of the Iso-Dalt system. ANL-BIM-79-2. Argonne National Laboratory 9700 South Cass Avenue, Argonne, Illinois 60409Google Scholar
  4. Black JA, Dixon GH (1968) Amino acid sequence of alpha chains of human haptoglobins. Nature 218:736–741CrossRefGoogle Scholar
  5. Connell GE, Dixon GH, Smithies O (1962) Subdivision of the three common haptoglobin types based on “hidden” differences. Nature 193:505–506CrossRefGoogle Scholar
  6. Connell GE, Smithies O, Dixon GH (1966) Gene action in the human haptoglobins. II. Isolation and physical characterization of alpha polypeptide chains. J Mol Biol 21:225–229CrossRefGoogle Scholar
  7. Hawkes R, Niday E, Gordon J (1982) A dot-immunobinding assay for monoclonal and other antibodies. Anal Biochem 119:142–147CrossRefGoogle Scholar
  8. Javid J (1967) Haptoglobin 2-1 Bellevue, a haptoglobin β-chain mutant. Proc Natl Acad Sci USA 57:920–924CrossRefGoogle Scholar
  9. Kurosky A, Barnett DR, Lee T-H, Touchstone B, Hay RF, Arnott MS, Bowman BH, Fitch WM (1980) Covalent structure of human haptoglobin: A serine protease homolog. Proc Natl Acad Sci USA 77:3388–3392CrossRefGoogle Scholar
  10. Nance WE, Smithies O (1963) New haptoglobin alleles: A prediction confirmed. Nature 198:869–870CrossRefGoogle Scholar
  11. Olaisen B, Mevåg B, Teisberg P, Gedde-Dahl T Jr, Helland R, Siverts A, Jonassen R, Hjalmarsson K, Swan T (1981) Plasma protein polymorphisms in two-dimensional electrofocusing/SDS electrophoresis. Proceedings. 9. International Congress of the Society for Forensic Haemogenetics, Bern 29.9–3.10.1981, pp 543–555Google Scholar
  12. Pastewka JV, Reed RA, Ness AT, Peacock AC (1973) An improved haptoglobin subtyping procedure using polyacrylamide gel electrophoresis. Anal Biochem 51:152–162CrossRefGoogle Scholar
  13. Shibata K, Constans J, Vian M, Matsumoto H (1982) Polymorphism of the haptoglobin peptides by isoelectric focusing electrophoresis and isoelectric point determinations. Hum Genet 61:210–214CrossRefGoogle Scholar
  14. Smithies O (1955) Zone electrophoresis in starch gels: Group variations in the serum proteins of normal human adults. Biochem J 61:629–641PubMedPubMedCentralGoogle Scholar
  15. Smithies O, Walker NF (1955) Genetic control of some serum proteins in normal humans. Nature 176:1265–1266CrossRefGoogle Scholar
  16. Smithies O, Walker NF (1956) Notation for serum-protein groups and genes controlling their inheritance. Nature 178:694–695CrossRefGoogle Scholar
  17. Smithies O, Connell GE, Dixon GH (1962a) Inheritance of haptoglobin subtypes. Am J Hum Genet 14:14–21PubMedPubMedCentralGoogle Scholar
  18. Smithies O, Connell GE, Dixon GH (1962b) Chromosomal rearrangements and the evolution of haptoglobin genes. Nature 196:232–236CrossRefGoogle Scholar
  19. Teige B, Mevåg B, Olaisen B, Pedersen L (1983) Haptoglobin subtyping in routine paternity cases—presentation of a promising new method. Proceedings, 10. International Congress of the Society for Forensic Haemogenetics, München, 11.–15. 10. 1983. pp 373–379Google Scholar
  20. Thymann M (1977) Improved haptoglobin subtyping demonstrating ά2 polymorphism in the Danish population. Proceedings. 7. International Congress of the Society for Forensic Haemogenetics, Hamburg 25.–29. 9. 1977, pp 187–192Google Scholar
  21. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354CrossRefGoogle Scholar
  22. Von Weerts G, Nix W, Deicher H (1966) Isolierung und nähere Charakterisierung eines neuen Haptoglobins: Hp-Marburg. Blut 12:65–77CrossRefGoogle Scholar
  23. Whitehouse DB, Putt W (1983) Immunological detection of the sixth complement component (C6) following flat bed polyacrylamide gel isoelectric focusing and electrophoretic transfer to nitrocellulose filters. Ann Hum Genet 47:1–8CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1985

Authors and Affiliations

  • Brita Teige
    • 1
  • Bjørnar Olaisen
    • 1
  • Lilian Pedersen
    • 1
  1. 1.Institute of Forensic MedicineUniversity of Oslo, RikshospitaletOslo 1Norway

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