In-process monitoring of protein purification with thin film silicon sensor technology.
We have developed a rapid and sensitive thin film assay for in-process monitoring of target protein purification. This novel biosensor method provides rapid (5-min) visual evaluation of column purification fractions. The method can be used to monitor the efficiency of purification and potential loss of protein if the column binding capacity is exceeded. The eluted fractions containing the highest yield of target protein can be quickly identified, pooled, and processed. This convenient platform, known as the SILAS product, is a thin-film detection technology in which specific molecular interactions are transduced into visible color changes based on changes in the optical thickness of layers on a silicon surface. The results are interpreted without instrumentation. Proteins eluted from a purification column are adsorbed to the assay surface, and the ligand of interest (target) can be identified with specific binding reagents. Here we demonstrate two protein purification applications for the SILAS technology product: monitoring antibody elution from a Protein G column and evaluating the efficiency of purification of a glutathione-S-transferase (GST)-tagged recombinant protein through each step of the purification process.
Ettinger
Anna
A
Thermo BioStar, Boulder, CO 80301, USA. a_ettinger@thermobiostar.com
Ostroff
Rachel
R
Brien
Jeanne
J
Polisky
Barry
B
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D015374:Biosensing Techniques / Q000379:methods*; D009344:Neisseria gonorrhoeae / Q000737:chemistry; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification*; D012825:Silicon
D000906:Antibodies; D011993:Recombinant Fusion Proteins; D012825:Silicon