International Summer School PROGRAM - Laboratoire d'Infochimie ...

2
ROLE OF SPACER IN THE PHASE-TO-PHASE LAYER
OF POLYMERIC MICROSPHERES AT CONSTRUCTING OF
BIOTEST-SYSTEMS
1 Fedorova O.V., 1 Zayarnyuk N.L., 2 Stanishevskiy Ya.M.,
2 Gritskova I.A., 1 Novikov V.P.
1
National University “Lviv Politechnic”
79013, Lviv, Bandery str., 12, e-mail: vnovikov@polynet.lviv.ua
Moscow State Academy of Fine Chemical Technology n. of M.V. Lomonosov
When constructing biotest-systems on viral and infectious diseases specific
immunoglobulins G (Ig G) have been used as bioligands. The immobilization of these
immunoglobulins on the surface of functional polymeric microspheres proposed to
through spacer, protein A (S. aureus). The choice of spaser is conditioned by its
capability of the affine interaction with Fc - fragment of molecule. In this case Fab -
fragment of Ig G provides more effective availability of binding to antigen’s
determinants with potential causative agent of a disease.
Optimal concentrations of protein A (0,05 mg/ml) on the surfaces of polymeric
microspheres and minimum dilutions of serums, at which particles of conjugates
"polymeric microsphere - protein A" spontaneously aggregate, have been determined.
The orientation of Ig G molecule with hydrophilic Fab-fragment into the water
phase results in lyophilization of a surface of a particle, which is the condition of the
development of structural - mechanical factor by Rebinder. The creation of such type of
diagnostic biotest-systems doesn’t require a preliminary selection and purification of
specific molecules of Ig G, which are contained in γ- globulin’s fraction required.
Stability of the conjugate "polymeric microsphere (PM) - protein A" allows using it
more than 6 months.
Biotest-systems of different specificity for the following diseases have been
developed using highly titratable serums and universal conjugate "PM - protein A":
yersiniosis (Y. enterocolitica); salmonellosis (S. pullorum); leptospirosis (L. conicula);
for the chicken infectious bronchitis virus stamm Н120.