An AAV viral gene vector must be purified from an anchoragedependent cell line. Repeat Exercise 1.21 to develop a purification process for this vector.

Data From Exercise 1.21

Recombinant protein production from E. coli resulted in the first products from biotechnology. 

(a) List the primary structures and components of E. coli that must be removed from a fermentation broth to purify a heterologous protein product (one that differs from any protein normally found in the organism in question) expressed for pharmaceutical use. 

(b) Identify a sequence of steps to purify a conjugate heterologous protein (a compound comprised of a protein molecule and an attached non-protein prosthetic group such as a carbohydrate)
that remained soluble in cell paste.

(c) Identify a separation operation for each step in the process and list one alternative for each step. 

(d) Summarize important considerations in establishing operating procedures to preserve the activity of the protein. 

(e) Suppose net yield in each step in your process was 80%. Determine the overall yield of the process and the scale of operation required to produce 100 kg per year of the protein at a titer of 1 g/L.