Apparatus and methods for fractionation of biological products
A technology of biological products and preparations, applied in the field of polynucleotide and virus particle devices, and protein purification, which can solve the problems of impaired productivity in the purification process
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Embodiment 1
[0202] Example 1. Extraction of chromatin from an IgG-containing cell culture harvest containing 259218 ppm host protein contaminants and 21% aggregates. Chromatin was extracted as follows: add 1% allantoin, 0.4% octanoic acid, pH 5.2, incubate for 2 hours, then add electropositive metal affinity particles ((TREN 40high, Bio-Works) to a volume ratio of 4%, and mix and incubate An additional 4 hours, then solids were removed by passing the formulation through a depth filter (Sartorius PC1). Host protein was reduced to 308 ppm and aggregates were reduced to less than 0.05%. The prototype device was configured with a single tangential flow filtration subunit and a single adsorber unit. The tangential flow filtration subunit is equipped with a polyethersulfone (PES) membrane with an average pore size (Millipore) corresponding to a hypothetical globulin with a hydrodynamic diameter of 50 kDa. The adsorption subunit is a strong anion exchange monolithic column (CIM QA , BIA Separati...
Embodiment 2
[0203] Example 2. The harvest of extracted chromatin from Example 1 was applied to a prototype device with the same tangential flow filtration subunit as in Example 1 but with the adsorption subunit replaced by a Sartobind phenyl membrane adsorber (Sartorius). The conditioned harvest was concentrated to approximately 20 mg / mL with the adsorption subunit online. The phenyl adsorbent subunit was brought online and the buffer was exchanged to 50 mM HEPES, 1.7M NaCl, pH 7.0. When the buffer flowing through the system reaches the same pH and conductivity as the input buffer, flush the phenyl sorbent subunit lines with clean 50 mM HEPES, 1.7M NaCl, pH 7.0 and take the sorbent unit offline. IgG was collected and analyzed. Aggregates were reduced to less than 0.01% and host proteins were reduced to 9 ppm.
Embodiment 3
[0204] Example 3. Application of the harvest of extracted chromatin from Example 1 to a Sartobind phenyl membrane adsorber (Sartorius) format with the same tangential flow filtration subunit and the same adsorption subunit (QA monolithic column) The prototype device of the additional adsorption subunit. The conditioned harvest was concentrated to approximately 20 mg / mL with the adsorption unit not online. The phenyl adsorbent subunit was brought online and the buffer was exchanged to 50 mM HEPES, 1.7M NaCl, pH 7.0. When the buffer flowing through the system reaches the same pH and conductivity as the input buffer, flush the phenyl sorbent subunit tubing and take it offline, bring the QA monolithic column sorbent subunit online and simultaneously bring the phenyl sorbent subunit tubing Switch to offline. The buffer was exchanged to 50 mM Tris, pH 8.0. When the buffer flowing through the system reaches the same pH and conductivity as the input buffer, flush the QA monolithic ...
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