A high-throughput, cost-effective approach to Protein A capture chromatography

Thermo Scientific offers protein A affinity resins for monoclonal antibody purification that demonstrates high dynamic binding performance at low flow rates maintaining superior binding capacity at high linear velocities where other chromatography media products cannot operate.


POROS Protein A chromatography offers multiple advantages:

Features:

  • 50 μm, rigid, polymeric resins designed for the purification of monoclonal antibodies
  • Resin backbone consists of crosslinked poly[styrene divinylbenzene] coated with a polyhydroxylated polymer
  • The coating is further derivatized by covalent immobilization of a recombinant Protein A

Advantages:

  • Optimized for very rapid mass transport
  • Offers superior capacity vs. flow rate performance with the physical and chemical stability of a rigid polymeric support
  • Deliver improved throughput without increasing column size

POROS MabCapture A Select (MCAS)

  • Newest addition to the Protein A high-throughput chromatography resin family
  • Combines past improvements made to our existing MabCapture A product with our own self-produced Protein A ligand
  • Lower price to help reduce the cost of clinical manufacturing

Figure 1. Performance as a function of flow rate and residence time. (A) The high dynamic binding capacity of POROS MabCapture A is maintained as linear flow rate increases over a 3-fold range. Only a 6 mg/mL, or 12%, decay is realized compared to agarose resin; in this example, a GE MabSelect™ medium that realizes a 21 mg/mL, or 45%, decay in DBC over a more limited flow rate range.
(B) POROS Protein A resins demonstrate higher dynamic binding capacity, mostly independently of the residence time attained. Operational flow rate and column bed heights do not need to be optimized to allow diffusion of the IgG to the rPA-coated surface of the chromatography bead. Capture can be efficient and robust. This allows faster processing to increase cycles on a smaller column and to better utilize column lifetime and increase cost savings or reduce process times in general.


Figure 2. High pH stability and column lifetime. The capacity of the resin is only slightly decreased after exposure to 0.1 N NaOH for 30 minutes per cycle at 500 cm/hr, for 50 sanitization cycles. The data indicate that the reuse capability of the resin is quite robust. Since the protein preparations used for this study contained proteases, it is difficult to determine if the hydroxide is causing the stability to decrease or if this is due to the proteases in the load. The data include both modes of degradation as a worst-case study. This is also considered a conservative study, since most processes do not utilize a hydroxide step after every cycle.

Figure 3. rPA leachate as a function of cycle number. It is important to understand the effect that higher binding capacity may have on rPA leachate levels. The rPA is covalently attached to the POROS base bead. This graph indicates that rPA leachate levels are still in the low-ppm range and decrease and plateau as typically seen with rPA resins. POROS Protein A resins result in low leached Protein A concentrations and plateau with use as expected. After 5 cycles of reuse, MCA Select and MCA leachate levels are ≤10 ppm (10 ng/mg IgG) or lower.

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