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P. G. Sveshnikov, T. A. Yagudin, E. V. Morozkina, E. V. Klyachko, S. S. Zatsepin, S. V. Benevolensky, O. B. Shemchukova, L. P. Pozdnyakova, O. N. Solopova
Production
of the humanized Fab fragment of the neutralizing antibody against rabies virus
Abstract
The aim of the investigation is to produce functional humanized Fab
fragment of the neutralizing antibody against rabies virus – a prototype of a
therapeutic drug, capable of substituting the horse and the donor anti-rabies
immunoglobulin. There was performed cloning and sequencing of variable light
and heavy chains fragments of a high affinity neutralizing antibody against
rabies virus. There was made a substitution of constant mouse fragments for
human ones and a production of humanized Fab fragments in the yeast expression
system. There were also investigated the immunochemical properties of the
obtained Fab fragments by EIA, electrophoresis in PAAG and Western blotting.
The humanized Fab fragment of the antibody against rabies virus, which exceeded
the initial antibody in terms of affinity, was produced. The high humanization
degree was proved with the used of the sera, specific towards human
immunoglobulins. The yield of the humanized Fab fragment made up 21 mg per 1 L of the culture medium, the absence of isolated light and heavy chains of the humanized Fab
fragment was testified.
Copyright (C) Chemistry Dept., Moscow State University, 2002
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