Expression of Bispecific Antibodies Using Cellular Display

The identification and selection of high-producing cell lines can be a resource- and time-consuming process. The screening effort can be simplified by assessing the potential for high expression (or a desired product quality attribute) of the individual cell directly in a mix of cells. Here, we describe protocols for the use of such a cellular display technology. Using alternate splicing, two mRNA constructs are generated at tunable ratios. The first mRNA codes for the secreted product, the second mRNA attaches a transmembrane domain to the antibody and directs it to the cellular membrane. The design of the basic construct as well as efficient ways to tune the strength of the cellular display is detailed in this chapter. Further, enrichment methods are provided enabling the flow cytometric sorting of a cell population based on the quantity of cellular display or on the product quality (heterodimerization level of a bispecific antibody).