Antibodies are indispensable tools for basic research as well as diagnostic and therapeutic applications. Consequently, the development of alternative manufacturing strategies which circumvent the hurdles connected to conventional antibody production technologies is of enormous interest. To address this issue, we demonstrate the synthesis of complex antibody formats, in particular immunoglobulin G (IgG) and single-chain variable fragment Fc fusion (scFv-Fc), in a microsome-containing cell-free system based on translationally active chinese hamster ovary (CHO) cell lysates. To mimic the environment for antibody folding and assembly present in living cells, antibody genes were fused to an endoplasmic reticulum (ER)-specific signal sequence. Signal-peptide induced translocation of antibody polypeptide chains into the lumen of ER microsomes was found to be the prerequisite for antibody chain assembly and functionality. In this context, we show the rapid synthesis of antibody molecules in different reaction formats, including batch and continuous-exchange cell-free (CECF) reactions, depending on the amount of protein needed for further analysis. In addition, we demonstrate site-specific and residue-specific labeling of antibodies with fluorescent non-canonical amino acids. In summary, our study describes a novel antibody production platform which combines the highly efficient mammalian protein folding machinery of CHO cells with the benefits of cell-free protein synthesis.
Abstract. In the article the authors consider a cognitive management method of the investment-building complex in the crisis conditions. The factors influencing the choice of an investment strategy are studied, the basic lines of the activity in the field of crisis-management from a position of mathematical modelling are defined. The general approach to decisionmaking on investment in real assets on the basis of the discrete systems based on the optimum control theory is offered. With the use of a discrete maximum principle the task in view of the decision is found. The numerical algorithm to define the optimum control is formulated by investments. Analytical decisions for the case of constant profitability of the basic means are obtained.
Antibodies are indispensable tools for therapy and diagnostics. Here, we present a novel technique that allows for an accelerated synthesis of functionally active antibodies. This is achieved by using a cell-free translation system based on mammalian cells.
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