BIOENGINEERING OF RECOMBINANT PROTEINS

Abstract

The work is devoted to substantiating the peculiarities of technological schemes, processes, and influencing factors in the production of recombinant proteins. It is noted that modern recombinant DNA technologies involve the production of monoclonal antibodies and certain basic medicinal substances through bioreactions using mammalian cells, yeast, or bacteria. According to recombinant technology, the required gene is isolated from one organism and inserted into a small fragment of DNA, which serves as a carrier, known as a vector. It is noted that an effective condition for the reproduction of recombinant DNA is the presence of a similar or unrelated host/recipient cell. Considering that, as a result of the bioprocess, the recombinant protein expressed is found in the biomass at low concentrations and is highly diluted, the separation of biomass systems is a crucial component in recombinant protein biotechnology. Separation can be achieved through filtration, sedimentation, separation, centrifugation, and other methods. The paper presents a generalised scheme for processing recombinant proteins. The technological specificity of the processes and equipment in the production of recombinant protein is substantiated–separation, centrifugation, etc. The paper presents a generalised scheme for processing recombinant proteins. The technological specifics of the processes and equipment used in the production of recombinant proteins are substantiated, and the factors that ensure effective bioprocessing in this process are identified. It has been proven that the critical stages of protein concentration are filtration, separation, centrifugation, followed by ultrafiltration and diafiltration to obtain a concentrated protein solution in a buffer solution. Purification of the concentrated protein is performed using ion-exchange or affinity chromatography. Final sterile filtration using microfiltration ensures the final product is a recombinant protein. The article establishes and substantiates the role of productivity factors in the stages of separation and concentration of primary protein, including protein yield, degree of solid phase separation, and centrifuge productivity.