Abstract

ANALYSIS OF THE FUNCTIONAL ACTIVITY OF THE ISOTYPES C4A AND C4B OF THE HUMAN COMPLEMENT SYSTEM IN A HYBRID MICROPANEL

Vladimir M. Lakhtin*, Mikhail V. Lakhtin, Vladimir A. Aleshkin, Alexey M. Vorobiev

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Abstract

Variants for analyzing the functional activity of the C4A and C4B isotypes of the C4 component of the human complement system in a hybrid flat-bottomed polystyrene 96-well micropanel are proposed and described. The analysis allows simultaneous determination of the functional activity of both isotypes under identical conditions of temporary assembly of components along the classical pathway of the human complement system. The choice is justified and the procedures for obtaining laboratory preparations of reagents for functional analysis of isotypes are described. Functional analysis options are promising for rapid assessment of both C4 and its isotype deficiencies in pathological sera. Results of the hybrid micropanel using were in accordance to the independent analysis of isotypes separated by isoelectrofocusing in a polyacrylamide gel plate and electroblotted on a membrane based on optimized chemiluminescence of blots in a live imagination. Hybrid micropanel procedures are promising in modeling and developing biand multitargeted disease control technologies, taking into account the extensive communication intermolecular and intercellular potential of the complement system in a single protective system of an individual's body.

Keywords: C4A, C4B, C4, complement system, serum, functional analysis, hybrid micropanel, diseases.