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Volume 89, Issue 1

Studies on the Purification of Chlamydial Agents Grown in Yolk Sacs of Embryonated Eggs Using Disulphide-linked Immunosorbents and Enzymes Free

Abstract

SUMMARY: Immunosorbents were derived from avid and non-avid sera raised in rabbits to multiple or single injections of chlamydiae passaged once or three times in HeLa cells after routine passage in eggs. Egg-derived suspensions of chlamydiae required pretreatment before application to immunosorbent columns; this was most conveniently done by fractionation on Sepharose 4B. Immunosorbents derived from avid serum had greater capacity than those from non-avid sera. However, organisms were desorbed in low yield from an avid immunosorbent, but in higher yields from non-avid immunosorbents. Even under the best conditions, the immunosorbents yielded suspensions of organisms still contaminated with egg antigens.

Partially purified suspensions of chlamydiae were also treated with phospholipase C to yield suspensions of high purity.

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© Society for General Microbiology, 1975
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1975-07-01
2024-04-25
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References

  1. Bartlett G. R. 1959; Phosphorus assay in column chromatography. Journal of Biological Chemistry 234:466–468
     [Google Scholar]
  2. Bird R. A., Low M. G., Stephen J. 1974; Immunopurification of phospholipase C (α-toxin) from Clostridium perfringens. FEBS Letters 44:279–281
     [Google Scholar]
  3. Birkbeck T. H., Stephen J. 1970; Specific removal of host-cell or vaccinia-virus antigens from extracts of infected cells by polyvalent disulphide-linked immunosorbents. Journal of General Virology 8:133–142
     [Google Scholar]
  4. Birkbeck T. H., Stephen J. 1971; Immunochemical isolation of vaccinia virus antigens. Immunochemistry 8:1029–1039
     [Google Scholar]
  5. Bligh E. G., Dyer W. J. 1959; A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology 37:911–917
     [Google Scholar]
  6. Chidlow J. W., Stephen J., Smith H. 1970; Further studies on the specific and quantitative aspects of antigen adsorption by disulphide-linked antibody immunosorbents. Immunochemistry 7:505–514
     [Google Scholar]
  7. Collier L. H. 1972; Some aspects of trachoma control and provisional estimates of the cost of vaccine production. Israel Journal of Medical Sciences 8:1114–1123
     [Google Scholar]
  8. Colón J. I., Moulder J. W. 1958; Folic acid in purified preparations of members of the psittacosis group of micro-organisms. Journal of Infectious Diseases 103:109–119
     [Google Scholar]
  9. Crook N. E., Chidlow J. W., Stephen J., Smith H. 1972a; Improved methods for the production of disulphide-linked antibody immunsorbents. Immunochemistry 9:749–752
     [Google Scholar]
  10. Crook N. E., Stephen J., Smith H. 1972b; Recovery of bound homologous protein antigens from disulphide-linked immunsorbents at neutral pH. Immunochemistry 9:945–960
     [Google Scholar]
  11. Crowle A. J. 1958; A simplified micro double diffusion agar precipitin technique. Journal of Laboratory and Clinical Medicine 52:784–787
     [Google Scholar]
  12. Detels R., Wang S. P., Grayston J. T. 1968; Febrile and local reactions to two highly concentrated trachoma vaccines with and without adjuvant. Revue internationale du trachome 45:331–340
     [Google Scholar]
  13. Dulbecco R., Vogt M. 1954; Plaque formation and isolation of pure lines with poliomyelitis virus. Journal of Experimental Medicine 99:167–182
     [Google Scholar]
  14. Garrett A. J., Harrison M. J., Manire G. P. 1974; A search for the bacterial mucopeptide component, muramic acid, in Chlamydia. Journal of General Microbiology 80:315–318
     [Google Scholar]
  15. Gogolak F. M., Ross M. R. 1955; The properties and chemical nature of the psittacosis virus haemagglutinin. Virology 1:474–496
     [Google Scholar]
  16. Hanna L., Thygeson P., Jawetz E., Dawson C. 1959; Elementary body virus isolated from clinical trachoma in California. Science; New York: 1301339–1340
     [Google Scholar]
  17. Howard L., Orenstein N. S., King N. W. 1974; Purification on Renografin density gradients of Chlamydia trachomatis grown in the yolk sac of eggs. Applied Microbiology 27:102–106
     [Google Scholar]
  18. Hughes M., Thomson R. O., Knight P., Stephen J. 1974; The immunopurification of tetanus toxin. Journal of Applied Bacteriology 37:603–621
     [Google Scholar]
  19. Jenkin H. M. 1960; Preparation and properties of cell walls of the agent of meningopneumonitis. Journal of Bacteriology 80:639–647
     [Google Scholar]
  20. Jones B. R. 1961; Trachoma and allied infections, opening paper. Transactions of the Ophthalmological Society of the United Kingdom 81:367–378
     [Google Scholar]
  21. Katzenelson E., Bernkopf H. 1965; Serological differentiation of trachoma strains and other agents of the psittacosis-lymphogranuloma venereum-trachoma group with the aid of the direct fluorescent antibody method. Journal of Immunology 94:467–474
     [Google Scholar]
  22. Katzenelson E., Bernkopf H. 1967; Studies of PLT agents with the aid of the agar immune diffusion technique. American Journal of Ophthalmology 63:1483–1487
     [Google Scholar]
  23. Kuo C. C., Kenny G. E., Wang S. P. 1971; Trachoma and psittacosis antigens in agar gel double immunodiffusion. In Trachoma and Related Disorders Caused by Chlamydial Agents. Proceedings of a Symposium held in Boston Massachusetts pp. 113–123 Nichols R. L. Edited by Amsterdam: Excerpta Medica.;
     [Google Scholar]
  24. Miller G. L., Golder R. H. 1950; Buffers of pH 2 to 12 for use in electrophoresis. Archives of Biochemistry 29:420–423
     [Google Scholar]
  25. Moulder J. W., Weiss E. 1951; Purification and properties of the agent of feline pneumonitis. Journal of Infectious Diseases 88:56–67
     [Google Scholar]
  26. Murray E. S. 1964; Guinea pig inclusion conjunctivitis virus. I. Isolation and identification as a member of the psittacosis-lymphogranuloma-trachoma group. Journal of Infectious Diseases 114:1–12
     [Google Scholar]
  27. Nicklin M. G., Stephen J. 1973; Solubilities of protein-antigen/rabbit-antibody complexes as a measure of serum avidity. Immunochemistry 10:717–725
     [Google Scholar]
  28. Nicklin M. G., Stephen J. 1974; Recovery of protein antigens from disulphide-linked immunosorbents derived from non-avid sera. Immunochemistry 11:35–40
     [Google Scholar]
  29. Ormsbee R. A., Weiss E. 1963; Trachoma agent: glucose utilisation by purified suspensions. Science; New York: 1421077–1078
     [Google Scholar]
  30. Provost P. J., Vickers J. H. 1972; Attempted immunization against trachoma infection in baboons. American Journal of Veterinary Research 33:599–605
     [Google Scholar]
  31. Reeve P., Taverne J. 1962; A simple method for total particle counts of trachoma and inclusion blennorrhoea viruses. Nature: London; 195:923–924
     [Google Scholar]
  32. Reeve P., Taverne J. 1967; Strain differences in the behaviour of TRIG agents in cell cultures. American Journal of Ophthalmology 63:1167–1173
     [Google Scholar]
  33. Ross M. R., Gogolak F. M. 1957; The antigenic structure of psittacosis and feline pneumonitis viruses. I. Isolation of complement-fixing antigens with group and species specificity. Virology 3:343–364
     [Google Scholar]
  34. Sarov I., Becker Y. 1968; RNA in the elementary bodies of trachoma agent. Nature: London; 217849–852
     [Google Scholar]
  35. Smith H., Tozer B. T., Gallop R. C., Scanes F. S. 1962; Separation of antigens by immunological specificity. I. Method for separating individual antigen-antibody complexes from mixed antigens and antibodies. Biochemical Journal 84:74–80
     [Google Scholar]
  36. Sowa S., Sowa J., Collier L. H., Blyth W. A. 1969; Trachoma vaccine field trials in the Gambia. Journal of Hygiene 67:699–717
     [Google Scholar]
  37. Stephen J., Gallop R. G. C., Smith H. 1966; Separation of antigens by immunological specificity. Use of disulphide-linked antibodies as immunosorbents. Biochemical Journal 101:717–720
     [Google Scholar]
  38. Sweet C., Stephen J., Smith H. 1974; Purification of influenza viruses using disulphide-linked immunosorbents derived from rabbit antibody. Immunochemistry 11:295–304
     [Google Scholar]
  39. Tamura A., Higashi N. 1963; Purification and chemical composition of meningopneumonitis virus. Virology 20:596–604
     [Google Scholar]
  40. Tang F. F., Chang H. L., Huang Y. T., Wang K. C. 1957; Studies on the etiology of trachoma with special reference to isolation of the virus in chick embryo. Chinese Medical Journal (Peking) 75:429–446
     [Google Scholar]
  41. Taverne J., Blyth W. A., Reeve P. 1964; Toxicity of the agents of trachoma and inclusion conjunctivitis. Journal of General Microbiology 37:271–275
     [Google Scholar]
  42. Vedros N. A. 1967; Species-specific antigens from trachoma and inclusion conjunctivitis (chlamydial) agents. Journal of Immunology 99:113–1186
     [Google Scholar]
  43. Wang S. P., Grayston J. T. 1967; A potency test for trachoma vaccine utilizing the mouse toxicity prevention test. American Journal of Ophthalmology 63:1443–1454
     [Google Scholar]
  44. Wang S. P., Kenny G. E., Grayston J. T. 1967; Characterisation of trachoma antigens protective against mouse toxicity. American Journal of Ophthalmology 63:1454–1461
     [Google Scholar]
  45. Watson D. H., Wildy P. 1969; The preparation of ‘monoprecipitin’ antisera to herpes virus specific antigens. Journal of General Virology 4:163–168
     [Google Scholar]
  46. Weiss E., Myers W. F., Dressler H. R., Chun-Hoon H. 1964; Glucose metabolism by agents of the psittacosis-trachoma group. Virology 22:551–562
     [Google Scholar]
  47. Weiss E., Schramek S., Wilson N. N., Newman L. W. 1970; Deoxyribonucleic acid heterogeneity between human and murine strains of Chlamydia trachomatis. Infection and Immunity 2:24–28
     [Google Scholar]
  48. Wood K. R., Gould E. A., Smith H. 1970; Purification of Semliki Forest virus using disulphide-linked immunosorbents derived from rabbit or fowl antibody. Journal of General Virology 6:257–265
     [Google Scholar]
  49. Wood K. R., Stephen J., Smith H. 1968; Specific adsorption of Semliki Forest virus on a disulphide-linked immimosorbent derived from rabbit antibody. Journal of General Virology 2:313–316
     [Google Scholar]
  50. Woodland R. M., Griffiths M. S., Pearce J. H. 1973; Adherence and specificity in chlamydial infections. Proceedings of the Society for General Microbiology 1:27
     [Google Scholar]
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Studies on the Purification of Chlamydial Agents Grown in Yolk Sacs of Embryonated Eggs Using Disulphide-linked Immunosorbents and Enzymes
Microbiology 89, 11 (1975); https://doi.org/10.1099/00221287-89-1-11
/content/journal/micro/10.1099/00221287-89-1-11
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