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Production and characteristics of monoclonal antibodies to the diphtheria toxin

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Abstract

Monoclonal antibodies to the diphtheria toxin were produced without cross reactivity with the thermolabile toxin (LT) from Escherichia coli; ricin; choleraic toxin; the SeA, SeB, SeE, SeI, and SeG toxins of staphylococcus; the lethal factor of the anthrax toxin; and the protective antigen of the anthrax toxin. A pair of antibodies for the quantitative determination of the diphtheria toxin in the sandwich variation of enzyme-linked immunosorbent assay (ELISA) was chosen. The determination limit of the toxin was 0.7 ng/ml in plate and 1.6 ng/ml in microchip ELISA. The presence of a secretion from the nasopharynx lavage did not decrease the sensitivity of the toxin determination by sandwich ELISA. The immunization of mice with the diphtheria toxin and with a conjugate of the diphtheria toxin with polystyrene microspheres demonstrated that the conjugate immunization resulted in the formation of hybridoma clones which produced antibodies only to the epitopes of the A fragment of the diphtheria toxin. The immunization with the native toxin caused the production of hybridoma clones which predominantly produced antibodies to the epitopes of the B fragment.

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Abbreviations

BSA:

bovine serum albumin

DT:

diphtheria toxin

DT-cPM:

the conjugate of the purified DT preparation with polystyrene microspheres

EIA:

enzyme immunoassay

TL:

thermolabile toxin

mAb:

monoclonal antibody

MIX1:

PBS with 1% polyvinyl alcohol, 1% polyvinylpyrrolidone, and BSA (10 mg/ml)

MIX2:

MIX1 with 0.1% Tween-20

PBS:

phosphate-salt solution (pH 7.4)

PBSP:

PBS with 1% polyvinyl alcohol

PBST:

PBS with 0.1% Tween-20

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Authors and Affiliations

  1. Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow, 117997, Russia

    T. I. Valyakina, O. E. Lakhtina, R. L. Komaleva, M. A. Simonova, L. V. Samokhvalova, N. S. Shoshina, N. A. Kalinina & E. V. Grishin

  2. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, ul. Vavilova 32, Moscow, 117991, Russia

    A. Yu. Rubina & M. A. Filippova

  3. Gamaleya Scientific Research Institute of Epidemiology and Microbiology, Russian Academy of Medical Sciences, ul. Gamaleya 18, Moscow, 123098, Russia

    Yu. V. Vertiev

Authors
  1. T. I. Valyakina
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  2. O. E. Lakhtina
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  3. R. L. Komaleva
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  4. M. A. Simonova
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  5. L. V. Samokhvalova
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  6. N. S. Shoshina
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  7. N. A. Kalinina
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  8. A. Yu. Rubina
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  10. Yu. V. Vertiev
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  11. E. V. Grishin
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Correspondence to T. I. Valyakina.

Additional information

Original Russian Text © T.I. Valyakina, O.E. Lakhtina, R.L. Komaleva, M.A. Simonova, L.V. Samokhvalova, N.S. Shoshina, N.A. Kalinina, A.Yu. Rubina, M.A. Filippova, Yu.V. Vertiev, E.V. Grishin, 2009, published in Bioorganicheskaya Khimiya, 2009, Vol. 35, No. 5, pp. 618–628.

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Valyakina, T.I., Lakhtina, O.E., Komaleva, R.L. et al. Production and characteristics of monoclonal antibodies to the diphtheria toxin. Russ J Bioorg Chem 35, 556–565 (2009). https://doi.org/10.1134/S1068162009050045

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  • DOI: https://doi.org/10.1134/S1068162009050045

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