Development of improved method of Isolation and Purification of human IgE

Автори

We aim at creating and evaluating the acceptability of improved methods of isolation of high purity human IgE suitable for use in highly sensitive immuno-analytical methods. To this end, we generalize physicochemical and biological properties of immunoglobulins of different classes and analyze key methodological techniques used for isolation and purification of human IgE (ion exchange chromatography on DEAE-cellulose, gel filtration on Sephadex G-200 and S-300 Sephacryl different options immunoaffinity chromatography). In addition, we justify circuit isolation and purification of human IgE based on own research data, as well as on other authors’ data. The technique is based on combination of the following methodological techniques: sequential removal of serum human IgG (by affinity chromatography on protein G), human IgA and IgM removal (using immunoaffinity chromatography based on sorbents with anti-IgA and anti-IgM monoclonal antibodies), allocation of human IgE by dual-gel filtration on Superdex 200. In addition, we perform the quality control of IgE by immunodiffusion for Ouhterloni and electrophoresis in polyacrylamide gel in reducing conditions. Using the proposed scheme, we obtain human IgE high purity. The human IgE output after all treatment phases is about 42% of the initial amount of immunoglobulin E in the serum.

Publication year: 
2013
Issue: 
3
УДК: 
УДК 577.27:57.083.33:543.54
С. 18—23. Іл. 4. Табл. 1. Бібліогр.: 22 назви.
References: 

1. Імунологія: Підручник / А.Ю. Вершигора, Є.У. Пастер, Д.В. Колибо та ін.; передм. С. Комісаренка; за заг. ред. Є.У. Пастер. — К.: Вища школа, 2005. — 599 с.
2. K. Ishizaka et al., “Physico-chemical properties of human reaginic antibody. IV. Presence of a unique immunoglobulin as a carrier of reaginic activity”, J. Immunol., vol. 97, no. 1, pp. 75 — 85, 1966.
3. D.R. Stanworth et al., “Specific inhibition of the Prausnitz Kьstner reaction by an atypical human myeloma protein”, Lancet, vol. 290 (7511), pp. 330—332, 1967.
4. A.J. Bonnin et al., “Association of cigarette smoking with elevated serum IgE levels in Hispanic Puerto Rican men”, Ann. Allergy, vol. 67, pp. 609—611, 1991.
5. A. Miadonna and C. Zanussi, “IgE synthesis in aging. Enhancement of IgE production in subjects with autoantibodies”, Boll. 1st. Sieroter. Milan., vol. 58, pp. 75—80, 1979.
6. K.J. Erb, “Helminths, allergic disorders and IgE-mediated immune responses: where do we stand?”, Eur. J. Immunol., vol. 37, no. 5, pp. 1170—1173, 2007.
7. J. Duarte et al., “Total and functional parasite specific IgE responses in Plasmodium falciparum-infected patients exhibiting different clinical status”, Malar. J., vol. 6, pp. 1—13, 2007.
8. I.V. Nikolayenko et al., “Preparation of highly purified human IgG, IgM, and IgA for immunization and immunoanalysis”, Ukrainica Bioorganica Acta, vol. 2, no. 2, pp. 3—11, 2005.
9. Иммунологические методы / Пер. с нем., под ред. Г. Фримеля. — М.: Медицина, 1987. — 472 с.
10. E. Harlow and D. Lane, Antibodies. A laboratory manual. New York: Cold Spring Harbor, 1988, 726 p.
11. Иммунология: Практикум / Е.У. Пастер, В.В. Овод, В.К. Позур, Н.Е. Вихоть. — К.: Вища школа, 1989. — 304 с.
12. Шугалей И.В., Гарабаджиу А.В., Целинский И.В. Химия белка: Учебн. пособие. — СПб.: Проспект Науки, 2010. — 200 с.
13. R.G. Hamilton, The human IgG subclasses, C. Mohan, Ed. Baltimore: Johns Hopkins University School of Medicine, 2001, 64 p.
14. J. Hakimi et al., “The alpha subunit of the human IgE receptor (FcERI) is sufficient for high affinity IgE binding”, J. Biol. Chem., vol. 265, no. 36, pp. 22079— 22081, 1991.
15. J. Kleine-Tebbe et al., “Purification of immunoglobulin E (IgE) antibodies from sera with high IgE titers”, J. Immunol. Methods, vol. 179, no. 2, pp. 153—164, 1995.
16. U.R. Laemmli, “Cleavage of structural proteins during the assembly of the of bacteriophage T4”, Nature, vol. 227, pp. 680 685, 1970.
17. Галкін О.Ю. Одержання імуноафінного сорбенту та розробка методики специфічного виділення IgM людини // Наукові вісті НТУУ “КПІ”. — 2009. — № 2. — С. 98—102.
18. L. Kеgedal et al., “Chemical, physical and chromatographic properties of Superdex 75 prep grade and Superdex 200 prep grade gel filtration media”, J. Chromatogr., vol. 537, pp. 17—32, 1991.
19. Галкін О.Ю., Дуган О.М. Порівняння схем імунізації мишей лінії Balb/c для одержання моноклональних антитіл до IgА людини // Науковий вісник Нац. унту біоресурсів та природокористування України. — 2009. — Вип. 134, ч. 1. — С. 88—97.
20. V.S. Chowdary et al., “A study on serum IgE and eosinophils in respiratory allergy patients”, Indian J. Allergy Asthma Immunol., vol. 17, no. 1, pp. 21—24, 2003.
21. T.A.E. Platts-Mills, “The role of immunoglobin E in allergy and asthma”, Am. J. Respir. Crit. Care. Med., vol. 164, pp. S1—S5, 2001.
22. N. Novak and T. Bieber, “Allergic and non-allergic forms of atopic disease”, J. Allergy. Clin. Immunol., vol. 112, pp. 252—262, 2003.

References [transliteration]: 

1. Imunolohii͡a: Pidruchnyk / A.I͡u. Vershyhora, I͡e.U. Paster, D.V. Kolybo ta in.; peredm. S. Komisarenka; za zah. red. I͡e.U. Paster. – K.: Vyshcha shkola, 2005. – 599 s.
2. K. Ishizaka et al., “Physico-chemical properties of human reaginic antibody. IV. Presence of a unique immunoglobulin as a carrier of reaginic activity”, J. Immunol., vol. 97, no. 1, pp. 75 – 85, 1966.
3. D.R. Stanworth et al., “Specific inhibition of the Prausnitz-Küstner reaction by an atypical human myeloma protein”, Lancet, vol. 290 (7511), pp. 330–332, 1967.
4. A.J. Bonnin et al., “Association of cigarette smoking with elevated serum IgE levels in Hispanic Puerto Rican men”, Ann. Allergy, vol. 67, pp. 609–611, 1991.
5. A. Miadonna and C. Zanussi, “IgE synthesis in aging. Enhancement of IgE production in subjects with autoantibodies”, Boll. 1st. Sieroter. Milan., vol. 58, pp. 75–80, 1979.
6. K.J. Erb, “Helminths, allergic disorders and IgE-mediated immune responses: where do we stand?”, Eur. J. Immunol., vol. 37, no. 5, pp. 1170–1173, 2007.
7. J. Duarte et al., “Total and functional parasite specific IgE responses in Plasmodium falciparum-infected patients exhibiting different clinical status”, Malar. J., vol. 6, pp. 1–13, 2007.
8. I.V. Nikolayenko et al., “Preparation of highly purified human IgG, IgM, and IgA for immunization and immunoanalysis”, Ukrainica Bioorganica Acta, vol. 2, no. 2, pp. 3–11, 2005.
9. Immunologicheskie metody / Per. s nem., pod red. G.Frimeli͡a. – M.: Medit͡sina, 1987. – 472 s.
10. E. Harlow and D. Lane, Antibodies. A laboratory manual. New York: Cold Spring Harbor, 1988, 726 p.
11. Immunologii͡a: Praktikum / E.U. Paster, V.V. Ovod, V.K. Pozur, N.E. Vikhot'. – K.: Vyssh. shkola, 1989. – 304 s.
12. Shugaleĭ I.V., Garabadzhiu A.V., T͡Selinskiĭ I.V. Khimii͡a belka: Uchebn. posobie. – SPb.: Prospekt Nauki, 2010. – 200 s.
13. R.G. Hamilton, The human IgG subclasses, C. Mohan, Ed. Baltimore: Johns Hopkins University School of Medicine, 2001, 64 p.
14. J. Hakimi et al., “The alpha subunit of the human IgE receptor (FcERI) is sufficient for high affinity IgE binding”, J. Biol. Chem., vol. 265, no. 36, pp. 22079–22081, 1991.
15. J. Kleine-Tebbe et al., “Purification of immunoglobulin E (IgE) antibodies from sera with high IgE titers”, J. Immunol. Methods, vol. 179, no. 2, pp. 153–164, 1995.
16. U.R. Laemmli, “Cleavage of structural proteins during the assembly of the of bacteriophage T4”, Nature, vol. 227, pp. 680–685, 1970.
17. Halkin O.I͡u. Oderz͡hanni͡a imunoafinnoho sorbentu ta rozrobka metodyky spet͡syfichnoho vydilenni͡a IgM li͡udyny // Naukovi visti NTUU “KPI”. – 2009. – # 2. – S. 98–102.
18. L. Kågedal et al., “Chemical, physical and chromatographic properties of Superdex 75 prep grade and Superdex 200 prep grade gel filtration media”, J. Chromatogr., vol. 537, pp. 17–32, 1991.
19. Halkin O.I͡u., Duhan O.M. Porivni͡anni͡a skhem imunizat͡siï mysheĭ liniï Balb/c dli͡a oderz͡hanni͡a monoklonal′nykh antytil do IgA li͡udyny // Naukovyĭ visnyk Nat͡s. un-tu bioresursiv ta pryrodokorystuvanni͡a Ukraïny. – 2009. – Vyp. 134, ch. 1. – S. 88–97.
20. V.S. Chowdary et al., “A study on serum IgE and eosinophils in respiratory allergy patients”, Indian J. Allergy Asthma Immunol., vol. 17, no. 1, pp. 21–24, 2003.
21. T.A.E. Platts-Mills, “The role of immunoglobin E in allergy and asthma”, Am. J. Respir. Crit. Care. Med., vol. 164, pp. S1–S5, 2001.
22. N. Novak and T. Bieber, “Allergic and non-allergic forms of atopic disease”, J. Allergy. Clin. Immunol., vol. 112, pp. 252–262, 2003.

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