Patent ID: 7157562

Claim:
A process for producing recombinant anti-botulinum toxin antibody, comprising the steps of: (a) fermenting recombinant E. Coli cells in broth, said recombinant E. Coli cells comprising DNA encoding anti-botulinum toxin antibody (Fab) proteins having a polyhistidyl tail at an end, said DNA under transcriptional control of a promoter induced by isopropyl beta-D-thiogalactopyranoside (IPTG), said fermenting producing a solution of said cells; (b) concentrating said cells by removing said broth from said solution by diafiltration; (c) crushing said concentrated cells producing crushed cells, cell debris, and retentate mixture; wherein said crushing comprises lysing the cells by mechanical means; (d) separating said retentate mixture from said crushed cells and cell debris, wherein said separating comprises passing said retentate mixture, crushed cells, and cell debris through a tangential flow ultrafiltration membrane; (e) separating DNA and other protein impurities from said retentate mixture, wherein said separation comprises passing said retentate mixture through a tangential flow ultrafiltration membrane producing a permeate; (f) purifying said recombinant antibotulinum antibody (Fab) from said permeate, wherein said purifying step comprises affinity chromatography, wherein said polyhistidyl tail at an end of said Fab provides a ligand, adding said permeate to a Ni—NTA resin in a bottle and rocking said bottle for at least 2 hours, pouring the resin/permeate mixture into a Ni—NTA affinity chromatography column, washing said column with a sonication buffer until the absorbancy at 280 nm is at a constant minimum, and eluting said Fab with an imidazole gradient of 0 mM to 500 mM producing imidazole eluate; (g) separating said Fab from said imidazole eluate by diafiltration; (h) quick freezing said Fab, and (i) freeze-drying said frozen Fab, said freeze-drying comprising lyophilization for stabilizing said Fab, said Fab comprising a protein product in a final purified, stabilized form.