Mammalian Cell Protein Expression Protocol

It is designed for high yield transient expression and display of mammalian cell surface membrane proteins including g protein coupled receptors gpcrs in an aqueous soluble format.

Mammalian cell protein expression protocol.

This unit reviews the stages involved in protein production in mammalian cells using a stable expression approach. Recombinant protein expression in mammalian cells hek293 cho introduction to expression in mammalian cells. Bacterial expression of recombinant his tagged proteins is a common technique. Recombinant proteins such as monoclonal antibodies are produced in mammalian cell lines to introduce proper protein folding and post translational modifications which are essential for full biological activity.

Choice of cell type is discussed as is transfection of the host cells methods for selection and. Complex glycosylation and accurate protein folding tasks where e. Coli or other hosts often fail make this host highly attractive although culturing and transfection costs are still significant. Chinese hamster ovary cho cells which have been successfully used for the manufacture of fda approved therapeutic proteins and antibodies are prominent in stable expression stably transfected cho cell line is the most suitable mammalian host for high yield production of recombinant proteins.

However use of other systems such as sf9 insect cells or hela or cho mammalian cells for expres sion of recombinant proteins either intracellularly or secreted into the culture medium is increasing. All are meant to rapidly lyse the cells with an easy to follow protocol. 1 chiron corporation emeryville california usa. Cellytic products are a family of protein extraction reagents specifically formulated to lyse and extract cellular proteins based on the type of expression system.

The gibco expi293 membranepro expression system combines the benefits of the expi293 expression system and the gibco membranepro expression system. These mammalian systems can produce full length and functionally active proteins with the hela systems typically expressing higher protein yields than rabbit reticulocyte system. However pro kary otic expression systems fail to generate correctly folded functional forms of many eukaryotic proteins and hence a variety of eukaryotic based expression systems have been developed 1. In both the industrial and academic environments the use of recombinant proteins varies widely and with it the method of.

The use of mammalian cells for the expression of recombinant proteins particularly of secreted proteins gains widespread use. The cellytic family is compatible with a wide variety of protease inhibitors chelating agents and chaotropes. Many proteins can be expressed at high level in escherichia coli. These eukaryotic expression systems may allow more natural.