Биологические науки / 9.Биохимия и
биофизика
Galkin O.Yu.
National
Technical University of Ukraine “Kyiv Polytechnic Institute”, Ukraine
NEW MONOclonal
ANTIBODIES AGAINST human IgA and their approach in immunoassay
Different
strategies of diagnostics and approaches for the construction of ELISA test-kits for determination
of certain infection are exist now. However all this strategies mainly are
based on the exposure of humoral immune response to the infection agent.
Elaboration of these test-kits is not possible without the use of antispecies
immunoenzyme conjugates. Conjugates of monoclonal antibodies (MAbs) to human IgA
are included in ELISA kits for the diagnostics of chlamydiosis, urea- and
mycoplasmosis, helicobacteriosis, hepatitis A infection and others [1-5].
The aim
of this development was an obtaining of new MAbs to human IgA, suitable for the
use as antispecies immunoenzyme conjugates in serological diagnostics.
An
original set of 25 new hybridoma cell-line that produces MAbs to human IgA is
obtained. The deep study of biological properties of antibodies has been
conducted. Comparative evaluation of horseradish peroxidase (HRP) conjugates of
MAbs is conducted and possibility of their usage in ELISA test-kits has been studied.
Two of them has shown better diagnostic value then known analogue GA-112 (Sigma).
The possibility of the usage of this MAbs conjugates in ELISA test-kits for the
diagnostics of helicobacteriosis, urogenital chlamydiosis, ureaplasmosis, urogenital
and respiratory mycoplasmosis is proved.
The
analysis of literary data about charts immunizations which are used in
hybridoma technology has been conducted. Results of own works on the obtaining
of monoclonal antibodies to human IgA by the different charts of immunization
has been generalized and analyzed. Possibility of the use of short-term and
long-term charts of immunization for the obtaining of monoclonal antibodies to
human IgA has been studied.
The possibility
of the application of two schemes of immunization for obtaining of anti-IgA MAbs
has been studied. Taking into account essential diagnostic value of MAbs, time
of receiving of antibodies, absence of allergic reactions for mice the
short-term scheme of immunization was more suitable and adequate. The close
correlation between the diversity of epitop specificity of MAbs and scheme of
immunization has been found out. At local immune response for Balb/c mice one district
of the polypeptide chain of IgA molecule has been appeared immunodominant. This
fact is confirmed by near epitop specificity of MAbs. At long-term scheme immune response was formed on various
antigen determinants.
References.
1.
Kohler G., Milstein C. Continuous
cultures of fused cells secreting antibody of predefined specificity // Nature.
– 1975. – Vol. 256. – P. 495 – 497.
2.
Kincade P. W., Lee Sun, Watanabe T. Monoclonal rat antibodies to murine IgM
determinants // J. Immunol. Methods. – 1981. – Vol. 42. – P. 17 – 26.
3.
Goding J. Monoсlonal antibodies. Principles
and practice. – San Diego: Acedemic press, 1996. – 492 р.
4. Климович В. Б., Самойловим М. П.,
Крутецкая И. Ю. и др. Моноклональные антитела к подласам IgG человека:
получение и исследование специфичности // Иммунология. – 1998. – №3. – С. 27 –
31.
5. Галкін
О.Ю., Дуган О.М. Порівняння схем імунізації мишей лінії Balb/c для одержання
моноклональних антитіл до IgА людини // Науковий вісник Національного
університету біоресурсів та природокористування України. – 2009. – Вип. 134, ч.
1. – С.88-97.