Establishing and developing CHO cell lines for the commercial production of human anti-Rhesus D IgG

摘要

Hemolytic Disease of the Newborn (HDN) can be a life threatening condition in Rhesus D positive babies borm from Rhesus D negative mothers. Successful prophylaxis in mothers is commonly based on the use of plasma-derived anti-D immunoglobulin preparations, which are rapidly becoming a scarce resource. In consequence, the substitution of the human derived polyclonal anti-D is both a health and an ethical concern. Several groups are involved in the expression of immunoglobulins anti Rh D in different hosts, including fused EBV-transformed human lymphocytes with murine myeloma (only IgG1) (Olovnikova, Belkina et al. 1997) and EBV-transformed B- lymphoblastoid cell lines (IgG1 and IgG3 subclasses) (Kumpel 1997). Among all the eukaryotic expression systems, CHI cells are still the most popular cell host for the production of recombinant proteins as human therapeutic agents (Wurm 1997), because of safety considerations, as well as practical issues related to scale up. Hoping to provide an alternative method for pharmaceutical manufacturing, we present here various aspects such as expressing vectors, transfection technology and selection strategies as approaches for the establishment of suitable CHO cell lines. We also tried to address the time and material constraints of our multidisciplinary team by using simple (rollerbottle) and more complex (2-20 L bioreactor) systems for production of human anti-Rhesus D antibodies.