Automated high-throughput protein purification using an ÄKTApurifier and a CETAC autosampler.
As the pace of drug discovery accelerates there is an increased focus on screening larger numbers of protein therapeutic candidates to identify those that are functionally superior and to assess manufacturability earlier in the process. Although there have been advances toward high throughput (HT) cloning and expression, protein purification is still an area where improvements can be made to conventional techniques. Current methodologies for purification often involve a tradeoff between HT automation or capacity and quality. We present an ÄKTA combined with an autosampler, the ÄKTA-AS, which has the capability of purifying up to 240 samples in two chromatographic dimensions without the need for user intervention. The ÄKTA-AS has been shown to be reliable with sample volumes between 0.5 mL and 100 mL, and the innovative use of a uniquely configured loading valve ensures reliability by efficiently removing air from the system as well as preventing sample cross contamination. Incorporation of a sample pump flush minimizes sample loss and enables recoveries ranging from the low tens of micrograms to milligram quantities of protein. In addition, when used in an affinity capture-buffer exchange format the final samples are formulated in a buffer compatible with most assays without requirement of additional downstream processing. The system is designed to capture samples in 96-well microplate format allowing for seamless integration of downstream HT analytic processes such as microfluidic or HPLC analysis. Most notably, there is minimal operator intervention to operate this system, thereby increasing efficiency, sample consistency and reducing the risk of human error.
Therapeutic Discovery, Amgen, Inc., One Amgen Center, Thousand Oaks, CA 91320-1789, USA.
Daniel
D
Yoo
Therapeutic Discovery, Amgen, Inc., One Amgen Center, Thousand Oaks, CA 91320-1789, USA.
Justin
J
Provchy
Therapeutic Discovery, Amgen, Inc., One Amgen Center, Thousand Oaks, CA 91320-1789, USA.
Cynthia
C
Park
Therapeutic Discovery, Amgen, Inc., One Amgen Center, Thousand Oaks, CA 91320-1789, USA.
Craig
C
Schulz
Therapeutic Discovery, Amgen, Inc., One Amgen Center, Thousand Oaks, CA 91320-1789, USA. Electronic address: kennethw@amgen.com.
Kenneth
K
Walker
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D002846:Chromatography, Affinity / Q000379:methods; D006801:Humans; D006825:Hybridomas / Q000737:chemistry; D051379:Mice; D046208:Microchip Analytical Procedures / Q000379:methods; D011506:Proteins / Q000302:isolation & purification*; D012371:Robotics
D000906:Antibodies; D011506:Proteins
Automated chromatography; High throughput purification; Multi-dimensional chromatography; ÄKTA