Process for recovering nonsecreted substances produced by cells

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Enzymatic production of a protein or polypeptide

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435178, 435182, 435241, 435803, 424 93, 424DIG7, C12P 2100, C12N 1110, C12N 502, A01N 6300

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045827990

ABSTRACT:
Disclosed is a process for recovering nonsecreted substances produced by cells. The process eliminates some of the high molecular weight contaminants thereby simplifying the purification process. The cells are encapsulated within a semipermeable membrane having properties which permit rapid passage of the relatively low molecular weight substances of interest but retard or prevent passage of higher molecular weight contaminants. The encapsulated cells are suspended in a culture medium and undergo normal cell growth and mitosis. The encapsulated cell culture grows to substantially fill the capsules but not rupture them. The cell membrane is then lysed without disrupting the capsule membrane. The permeability of the capsule membrane is such that the substances of interest diffuse rapidly through the capsule membrane into the extracapsular fluid while the higher molecular weight contaminants and cell fragments are retained within the capsule. The process is particularly useful for obtaining low to moderate molecular weight substances produced by pyrogen-producing prokaryotic, genetically modified organisms because the crude product can be collected relatively free of high molecular weight pyrogenic contaminants.

REFERENCES:
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Alberts, B., D. Bray, J. Lewis, M. Raff, K. Roberts and J. Watson, 1983 Molecular Biology of the Cell, pp. 265-266, Garland Publishing Inc.
Biotechnology & Bioengineering, vol. XIX, p. 387; M. Kierstan & C. Bucke (1977).

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