Ukr.Biochem.J. 2019; Volume 91, Issue 3, May-Jun, pp. 90-98


New monoclonal antibodies to the Chlamydia trachomatis main outer membrane protein and their immunobiological properties

O. Yu. Galkin1,2, O. B. Besarab1, Yu. V. Gorshunov1, O. M. Ivanova3

1National Technical University of Ukraine “Igor Sikorsky Kyiv Polytechnic Institute”;
2Propharma Plant Ltd., Kyiv;
3Xema Ltd., Kyiv

Received: 18 July 2018; Accepted: 14 March 2019

One of the methods that have been widely used in the diagnosis of urogenital chlamydia is an enzyme-linked immunosorbent assay (ELISA), the use of which allows for differential diagnosis. In order to increase the efficiency of ELISA test kits production, for the kits for the diagnosis of urogenital chlamydia, based on the principle of indirect modification, following synthetic positive controls (PCs) can be used: a conjugate of IgM (IgA) normal immunoglobulins and monoclonal antibodies (McAbs) to C. trachomatis major outer membrane protein (MOMP). The goal of this work was to obtain high active and affinity McAbs to the C. trachomatis MOMP as well as the study of its immunobiological properties which are important for future biochemical approaches. The study was conducted using: polyclonal antibodies (PcAbs) to C. trachomatis; recombinant major outer membrane protein (MOMP) (191-354 a.r.; W4-W5); epitope mapping based on phage display technology. The original set from 16 clones of hybridomas, producers of McAbs to the C. trachomatis MOMP has been obtained. More than half of the tested McAbs (8 out of 14) were characterized by a rather high titer (≥1:800), and three of them had a titer of ≥1:1600. In general, the McAbs titer was correlated with the value of the affinity constant: McAbs with higher titles were characterized by a high value of the affinity constant. For McAbs with a titer of <1:800, the average Ka is 5.2×109 M-1, while for McAbs with a titer ≥1:800 – Ka = 10.7×109 M-1. Antigenic determinants of two McAbs 293F4 and 291F8 that actively competed with PcAbs are represented by two linear sequences of 320-325 a.r. and 326-330 a.r., respectively. The epitope, which interacts with McAb 296G2, is represented by a linear sequence of 347-352 a.r. McAb 296G2 did not show active competition with serum PcAbs. The resulting set of data allows selecting McAbs for use in PCs of the ELISA kit for the detection of IgA or IgM antibodies to C. trachomatis.

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  1. Mavrov GI, Scherbakova YuV, Chinov GP. The treatment of complicated chamydial infection with azithromycin (Sumamed) using in combination with pathogenetic therapy. Ukr J Dermatol Venerol Cosmetol. 2010; (3(38)): 123-127.
  2. Galkin OYu, Gorshunov YuV, Besarab OB, Ivanova OM. Development and characterization of highly informative ELISA for the detection of IgG and IgA antibodies to Сhlamydia trachomatis. Ukr Biochem J. 2018;90(3):70-83.  CrossRef
  3. Isakov VA, Kulyashova LB, Berezina LA, Svarval AV. Laboratory diagnostics of urogenital chlamydiosis. Part 2. Methods of diagnosis of chlamydial infection: an analytical review. Terra Medica. Labor Diagn. 2013;(1): 13-21.
  4. Galkin OY, Gorshunov YV, Besarab OB. Synthetic positive controls for ELISA test kits for detection of IgA and IgM antibodies to Chlamydia trachomatis. Visn Dnipropetr Univ. Ser Biol Med. 2015; 6(1): 3-8.  CrossRef
  5. Galkin OYu, Savchenko AA, Nikitina KI, Dugan OM. Obtaining and study of properties of new monoclonal antibodies against human IgE. Ukr Biokhim Zhurn. 2013 Sep-Oct;85(5):81-7. (In Ukrainian).  PubMed,  CrossRef
  6. Nikolaenko IV, Galkin AIu, Raevskaia GE, Kas’ianenko TV, Nereshchenko MI, Donskaia ES, Spivak NIa. Preparation of monoclonal antibodies to the Fc-fragment of human IgG and the use of their based immunoenzyme conjugates. Klin Lab Diagn. 2005 Nov;(11):8-11. (In Russian). PubMed
  7. Goding J. Monoсlonal Antibodies: Principles and Practice. San Diego: Academic Press, 1996. 492 p.
  8. Shirobokov VP, Kopanitsa LV, Nikolaenko IV, Lipatnikova KI, Sologub VK. Monoclonal antibodies used to differentiate between poliovirus types 1 and 3. Mikrobiol Zh. 2001 Nov-Dec;63(6):42-52. (In Russian). PubMed
  9. Klimovich VB, Samoilovich MP, Krutetskaya IY. Monoclonal antibodies to human IgG subclasses: obtaining and investigation of specificity. Immunol. 1998; (3): 27-31.
  10. Köhler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495-7. PubMed, CrossRef
  11. Lane D, Koprowski H. Molecular recognition and the future of monoclonal antibodies. Nature. 1982 Mar 18;296(5854):200-2. PubMed, CrossRef
  12. Friguet B, Chaffotte AF, Djavadi-Ohaniance L, Goldberg ME. Measurements of the true affinity constant in solution of antigen-antibody complexes by enzyme-linked immunosorbent assay. J Immunol Methods. 1985 Mar 18;77(2):305-19. PubMed, CrossRef
  13. Kim BB, Dikova EB, Sheller U, Dikov MM, Gavrilova EM, Egorov AM. Evaluation of dissociation constants of antigen-antibody complexes by ELISA. J Immunol Methods. 1990 Aug 7;131(2):213-22. PubMed, CrossRef
  14. Bobrovnik SA, Demchenko MA, Komisarenko SV. The effect of serum polyreactive immunoglobulins on antibody affinity determination. Ukr Biokhim Zhurn. 2010 Jan-Feb;82(1):62-9. (In Russian). PubMed
  15. Rechkina EA, Denisova GF, Masalova OV, Lideman LF, Denisov DA, Lesnova EI, Ataullakhanov RI, Gurianova SV, Kushch AA.  Epitope mapping of antigenic determinants of hepatitis C virus proteins by phage display. Mol Biol (Mosk). 2006 Mar-Apr;40(2):357-68. (In Russian).
    PubMed,   CrossRef
  16. Galkin OYu. Comparative characteristic of the methods of protein antigens epitope mapping. Ukr Biochem J. 2014 Jul-Aug;86(4):164-77. (In Ukrainian). PubMed, CrossRef
  17. Wang Y, Berg EA, Feng X, Shen L, Smith T, Costello CE, Zhang YX. Identification of surface-exposed components of MOMP of Chlamydia trachomatis serovar F. Protein Sci. 2006 Jan;15(1):122-34.  PubMed, PubMedCentral, CrossRef
  18. Borgoyakova MB, Ilichev AA. Mapping of the epitope on the ERBB-2 protein, recognized by the monoclonal antibody 4B8, using phage display technology. Siber Oncol J. 2013;(4(58)): 40-44. (In Russian).
  19. Maksyutov AZ, Bachinskii AG, Bazhan SI, Ryzhikov EA, Maksyutov ZA. Exclusion of HIV epitopes shared with human proteins is prerequisite for designing safer AIDS vaccines. J Clin Virol. 2004 Dec;31 Suppl 1:S26-38. PubMed, CrossRef
  20. Amarasinghe S, Kathriarachchi H, Udagama P. Conserved regions of Plasmodium vivax potential vaccine candidate antigens in Sri Lanka: conscious in silico analysis of prospective conformational epitope regions. Asian Pac J Trop Med. 2014 Oct;7(10):832-40. PubMed,  CrossRef
  21. Baehr W, Zhang YX, Joseph T, Su H, Nano FE, Everett KD, Caldwell HD. Mapping antigenic domains expressed by Chlamydia trachomatis major outer membrane protein genes.  Proc Natl Acad Sci USA. 1988 Jun;85(11):4000-4. PubMed, PubMedCentral, CrossRef
  22. Batteiger BE. The major outer membrane protein of a single Chlamydia trachomatis serovar can possess more than one serovar-specific epitope. Infect Immun. 1996 Feb;64(2):542-7. PubMed PubMedCentral
  23. Stephens RS, Wagar EA, Schoolnik GK. High-resolution mapping of serovar-specific and common antigenic determinants of the major outer membrane protein of Chlamydia trachomatis. J Exp Med. 1988 Mar 1;167(3):817-31. PubMed, CrossRef
  24. Zhong GM, Reid RE, Brunham RC. Mapping antigenic sites on the major outer membrane protein of Chlamydia trachomatis with synthetic peptides. Infect Immun. 1990 May;58(5):1450-5. PubMed, PubMedCentral
  25. Savel’eva NV, Zagryadskaya YuE, Klimashevskaya SV, Puzyrev VF, Burkov AN, Obryadina AP, Ulanova TI. Scanning diagnostically significant antigenic regions of major Chlamydia trachomatis protein MOMP using series of overlapping recombinant proteins. Mol Genet Microbiol Virol. 2009; 24(3): 17-21. (In Russian). CrossRef

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