ResourcesCapture and purification of human serum albumin from human source plasma using Mimetic Blue® SA HL P6XL
Mimetic Blue® affinity chromatography adsorbents developed and manufactured exclusively by Astrea Bioseparations Ltd (‘Astrea’) comprise Mimetic Blue® ligands attached to beaded agarose supports. These adsorbents selectively bind albumin and albumin related proteins from a variety of sources including plasma, serum and recombinant cell cultures to yield highly purified biological products.
Mimetic Blue® SA HL P6XL has a high binding capacity for albumin and is intended for use as the initial capture and purification step. Mimetic Blue® SA HL P6XL is stable across a very wide pH range (pH 2.0 to pH 14.0), exhibits negligible ligand leakage by ELISA[1] and maintains chromatographic performance over 350 process cycles[1]. This adsorbent is used in regulated large scale capture applications.
Human plasma contains approximately 30-35 g/L of albumin in a complex mixture of proteins including IgG, fibrinogen and alpha 1-antitrypsin. Mimetic Blue® SA HL P6XL captures and purifies albumin from human plasma with high binding capacities and high purity.
In this application note we describe how human serum albumin is captured and purified from human source plasma using Mimetic Blue® SA HL P6XL.
Materials & Method
Human source plasma (untreated) was loaded onto equilibrated Mimetic Blue® SA HL P6XL packed to a bed height of 15 cm in a 1 cm diameter column. After loading, non-bound protein was removed by washing with equilibration buffer. The bound albumin was eluted using a buffer containing sodium octanoate. Mimetic Blue® SA HL P6XL is hydroxide stable and was cleaned with 0.5 M NaOH. The conditions are summarised in Table 1.
TABLE 1
Chromatography conditions for the capture and recovery of human serum albumin from human source plasma using Mimetic Blue® SA HL P6XL.
RESULTS & DISCUSSION
Mimetic Blue® SA HL P6XL selectively binds to the albumin in plasma, while other plasma proteins are not retained by the adsorbent and are present in the flow through fraction (Figures 1 and 2).
The resulting eluate has a purity of 96.1%, as determined by densitometry (Figure 2 and Table 2).
TABLE 2
Performance data for Mimetic Blue® SA HL P6XL determined by Nephelometry and densitometry.
*The flow through sample, measured by Nephelometry, was below the limit of detection. As a result, the Yield (%) result is presented as a less than value.
Mimetic Blue® SA HL P6XL has a dynamic binding capacity in excess of 20 g/L of adsorbent and high recovery, as determined by Nephelometry (Table 2).
CONCLUSIONS
Mimetic Blue® SA HL P6XL can be used to capture and purify human serum albumin from human source plasma with a dynamic binding capacity and recovery in excess of 20 g/L of adsorbent with a purity of ~96%. Mimetic Blue® SA HL P6XL has a dynamic binding capacity of in excess of 30 g/L of adsorbent for albumin in other feedstocks.
For further information please e-mail: sale@astrea-bio.com.
REFERENCES
[1] Mimetic Blue® Affinity Adsorbents Brochure (Issue: 060510).
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