Many of the drugs and other products that biotechnology companies produce are based on monoclonal antibodies. Biotechnicians who know how to work with and purify these materials are in demand.
Recently, the Northeastern Biomanufacturing Center and Collaborative (NBC2) made new curriculum modules available that include procedures for the upstream and downstream methods used to produce a specific anti-IL8 monoclonal antibody from CHO cells. This work was funded by a grant from the National Science Foundation’s Advanced Technology Education program (DUE #1501631).
Since antibodies play such a critical role in biotechnology, I thought it would good to write a short series on antibodies, their central role in the biotech industry, and the steps our students need to understand when it comes to antibody validation.
What are monoclonal antibodies?
One of the most important milestones in biotechnology happened in 1975 when César Milstein and Georges Köhler published a method for fusing B cells with myeloma cells to make cellular hybrids called “hybridomas.” These new hybrid cells inherit the ability to produce antibodies from their B cell parent and immortality from their cancer parent (the myeloma). When hybridoma cells are diluted and plated as single cells, they divide and produce clones. Since each clone contains the same rearranged antibody genes (more on this later) all the clones make the identical antibody. Since this clonal population of cells matkes the same antibody, the antibodies are described as monoclonal.
To produce large monoclonal antibodies in a large amount, the genes that encode the heavy and light chains are cloned, put into an expression vector, and transfected into mammalian cells. In the NBC2 curricula, the heavy and light chain genes that encode the anti-IL8 antibody were obtained from a mouse, put into an expression vector, and transfected into CHO cells for production. The CHO cells that are used to produce the antibody were originally obtained from a Chinese hamster’s ovary (CHO cells). Luckily, we can order the transfected cells from ATCC.
What is in the curriculum?
The NBC2 curriculum is very comprehensive. The curriculum modules include procedures for growing the CHO cells, harvesting the antibody, performing quality control procedures in addition to forms for detailed batch records, corresponding SOPs, and a list of the required equipment and reagents for production, and associated quality control procedures.
Biomanufacturing procedures are described as upstream and downstream to distinguish between the events that happen before harvest (upstream) and after harvest (downstream). The upstream processes consist of growing the cells, monitoring their condition, and harvesting the cells when they’ve reached the right point. The downstream processes involve purifying the desired material. Each phase of the procedure has it’s own methods and measurements for quality control.