Tag Archives: monoclonal antibodies

Effect of trifluoroethanol on antibodies binding properties

S. A. Bobrovnik*, M. O. Demchenko, S. V. Komisarenko

Department of Molecular Immunology, Palladin Institute of Biochemistry,
National Academy of Sciences of Ukraine, Kyiv;
*e-mail: s-bobrov@ukr.net

Received: 01 December 2022; Revised: 22 February 2023;
Accepted: 13 April 2023; Available on-line: 27 April 2023

The studies on the influence of organic co-solvents on the structure and function of antibodies are of key interest, especially in view of antibodies broad use as recognizing elements in different analytical systems. Here we studied the effect of co-solvent 2,2,2-trifluoroethanol (TFE) on the ability of anti-ovalbumin monoclonal antibodies to interact with its specific antigen. Antibody affinity to antigen and the rate constants of antibody binding to immobilized antigen were analyzed. Changes in antibody reactivity with incubation time which depended on TFE concentration and temperature were revealed. When treatment of antibodies with TFE was carried out at 0°C, we observed nonlinear, non-monotonous changes of antibody reactivity with initial fast decrease and substantial increase as incubation continued that may be related to the loss of antigen binding reactivity by some part of antibodies at the start but its restoration when the incubation proceeds.

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”;
e-mail: alexfbt@gmail.com;
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.

Inventive activity of the Department of Molecular Immunology of the Palladin Institute of Biochemistry of NAS of Ukraine

V. M. Danilova, R. P. Vynogradova, S. G. Torkhova

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: valdan@biochem.kiev.ua

The article is devoted to the inventive activity of the Department of Molecular Immunology of the Palladin Institute of Biochemistry of NAS of Ukraine in the context of the history of its inception, development and in the context of scholarly and organizational activities of Sergii Vasyl’ovych Komisarenko. This autumn marks 50th anniversary since young Sergii Komisarenko (now – Academician of NAS and NAMS of Ukraine, Dr. Biol. Sci., Professor) has joined the Palladin Institute of Biochemistry, has completed all stages of the academic carrier from PhD student to Head of the Institute. He is the first in Ukraine who started the new branch of research – molecular immunology, created a strong scientific school, which earned worldwide acclaim and made significant contribution to finding solutions to current problems in human health sciences. S.V. Komisarenko was among those, who were first in the USSR to use immunoenzyme and flow cytofluometric assays, hybridoma technology for producing monoclonal antibodies and immunochemical assay of proteins, which became the basis for development of highly sensitive and highly specific immunodiagnostic systems, which are of high necessity in medicine, veterinary, development of immunotechnologies, environment monitoring, etc.
Under his leadership the Department has made a series of important discoveries and developments including relating to antitumour immunotoxins, effects of low dose radiation on the immune system of Chernobyl liquidators, immunochemical structure of neurotoxin apamine, cytochrom c, fibrinogen and fibrin molecules at different stages of polymerization, diphtheria toxin and its receptor, tuberculosis causing micobacterium, roles of protease-activated receptors (PARs) and nicotinic acetylcholine receptors of lymphocytes, nature of polyreactive immunoglobulins (PRIGs), among other important scientific contributions.
S.V. Komisarenko and his colleagues also hold numerous (more than 80) author’s certificates and patents in Ukraine and USSR.

Inventive activity of the Departments of Protein Structure and Function, and Molecular Immunology of the Palladin Institute of Biochemistry of NAS of Ukraine. Part II. National breakthrough in the study and diagnostics of human hemostasis system

N. E. Lugovska

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: nlugovsk@mail.ru

The scientists of Protein Structure and Function, and Molecular Immunology Departments of the Palladin Institute of Biochemistry (NAS of Ukraine) under the supervision of member of NASU and NAMSU, prof. S. V. Komisarenko  and corresponding member of NASU prof. E. V. Lugovskoy have made the real breakthrough in the field of research of the mechanisms of fibrin polymerization and formation of fibrin framework of thrombi.  The immunodiagnostic test-systems for the evaluation of the risk of thrombus formation  were developed for the first time. Researches have obtained the monoclonal antibodies to fibrinogen, fibrin, D-dimer and their fragments. These monoclonal antibodies were used as molecular probes for the localization of newly detected fibrin polymerization sites. Obtained antibodies with high affinity interact with fibrinogen, D-dimer and soluble fibrin – main markers of the risk of thrombus formation. They were used for the development of the immunodiagnostic test-systems to quantify these markers in human blood plasma for the evaluation of the state  of haemostasis system, detection of prethrombotic states, disseminated intravascular coagulation, detection of thrombosis and monitoring of antithrombotic and fibrinolytic therapy. The successful trial of developed test-systems was carried out in clinics of Ukraine, and the State registration was obtained for the implementation of them into the clinical practice. Presented works were awarded State prize of Ukraine in Science and technology.

Obtaining and study of properties of new monoclonal antibodies against human IgE

O. Yu. Galkin1,2, A. A. Savchenko1, K. I. Nikitina1, O. M. Dugan1

1National Technical University of Ukraine “Kyiv Polytechnic Institute”;
2Hema Ltd., Kyiv, Ukraine;
e-mail: alexfbt@mail.ru

The original set of 12 clones of hybridomas, producers of monoclonal antibodies (mAbs) against human IgЕ, has been obtained. The study of biological properties of antibodies has been conducted: their specificity, affinity constant and titer in a cultural medium have been established. The obtained mAbs are directed to the two epitope regions on IgE molecule. The first group of mAbs relates to epitope region, represented by three epitopes (two of them overlap and one that does not overlap with others). The second epitope region is represented by only one epitope.

Laureates of the Palladin Prize of the National Academy of Sciences of Ukraine (2001, 2003 years)

V. M. Danilova, R. P. Vynogradova

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: valdan@biochem.kiev.ua

In 2001 L. L. Gromashevska (1922-2009), Head of Laboratory at L. V. Gromashevsky Institute of Epidemiology and Infectious Diseases, NAMS of Ukraine and A. S. Mikosha, Head of Laboratory at V. P. Komisarenko Institute of Endocrinology and Metabolism, NAMS of Ukraine were awarded the Palladin Prize of NAS of Ukraine for the series of works “The regulation of biochemical processes in norm and pathology“, which presents the results of the research on hormone and enzyme biochemistry, signaling pathways in cells under different physiological and pathological conditions.
In 2003 S. V. Komisarenko, E. V. Lougovskoy and I. M. Kolesnikov, Director and staff of Palladin Institute of Biochemistry, NAS of Ukraine, respectively, were awarded the Palladin Prize of NAS of Ukraine for the series of works “Immunochemical analysis of mechanisms of fibrin polymerization and fibrinolysis“. The paper presents an analysis of scientific activity and laureates’ biographical information.