Chemistry: molecular biology and microbiology – Micro-organism – per se ; compositions thereof; proces of...
Patent
1996-06-21
1999-06-22
Housel, James C.
Chemistry: molecular biology and microbiology
Micro-organism, per se ; compositions thereof; proces of...
209 31, 209511, 209552, 209576, 356 73, 424 93, 424493, 435177, 435178, C12N 100, B07C 502, G01N 2100, A01N 6300
Patent
active
059142626
DESCRIPTION:
BRIEF SUMMARY
SCOPE OF THE INVENTION
This invention relates to a novel method of microbial screening, and to a novel apparatus for carrying out the method.
BACKGROUND OF THE INVENTION
In the fields of microbiology, fermentation etc. it is frequently necessary to screen microbial cultures to distinguish between cells with different attributes or identify cells with similar attributes, for example viable and non-viable cells, or the level of productivity of a useful metabolite. One field in which such screening is particularly useful is in biotechnology, where microbial line development involving mutation of cells by mutagenic radiation or chemicals is carried out. After a microbial culture has been exposed to mutagenic influences it is necessary for the culture to be screened to detect mutations of interest. As the ratio of useful mutations to useless mutations is usually very low and the number of cells to be screened is usually very large it is desirable that fast and effective, ideally automated, processes are provided to carry out as much as possible of the screening.
Automated cell screening apparatus are known, but at present these can only screen relatively small numbers of cultures per year, and there is a need to improve on the rate of screening.
SUMMARY OF THE INVENTION
The inventors have developed a novel, improved microbial screening process and apparatus for carrying this process out.
According to a first aspect of this invention, a method of distinguishing between microorganisms having different attributes comprises the steps of: time sufficient for multiplication of viable cells within the bead; the bead for physical characteristics associated with the differing attributes of the cells within the bead.
DESCRIPTION OF THE DRAWINGS
An apparatus and method of this invention will now be described by way of example only with reference to the following figures:
FIG. 1 which shows an apparatus of the invention in an overall schematic arrangement.
FIG. 2 which shows an alternative construction of the manifold of the apparatus of FIG. 1.
FIGS. 3A-C which show alternative forms of particle directing means for use in the apparatus of FIG. 1.
FIGS. 4A-F which show alternative forms of particle directing means.
FIG. 5 which shows a further alternative form of particle directing means.
FIG. 6 which shows a further alternative form of particle directing means.
DETAILED DESCRIPTION OF THE INVENTION
The term "cell" used herein includes bacterial cells, animal or plant cells, fungal or bacterial spores (including conidia), microbial protoplasts and mycelial fragments. Example of suitable cells include spores of Penicillium chrysogenum and Streptomyces clavuligerus.
The method may be optionally followed, after further incubation, by the further step or steps of separating beads which contain cells possessing a selected attribute from beads which do not contain such cells, for example separating beads which contain multiplied viable cells from beads containing non-viable cells, or separating beads which contain one type of cell from beads which contain another type, or separating beads which contain cells which differ in their degree of viability and/or level of growth. This may then be followed by a yet further step of culturing and growing the viable cells on a larger scale, which may in turn be followed by a yet further step of examining and/or screening the grown viable cells for some useful characteristic. This may ultimately be followed by large scale fermentation.
The cells may be for example fungal or microbial spores. The method is particularly suited to screening for viable cells subsequent to exposure of the cells to a mutagenic influence, prior to subsequent screening for usefulness, e.g. secretion of a cell product such as an antibiotic.
Preferably a single cell is encapsulated within each bead.
Methods of encapsulation of single cells within polymeric beads are known. The polymeric bead is suitably an alginate polymer bead, in particular a calcium alginate polymer bead. One suitable method of encaps
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patent: 4175662 (1979-11-01), Zold
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Addicott John Kenneth
Arlington Stephen Adrian
Divers Moira Sloan
Drake David
MacMichael Donald Bruce Atherton
Housel James C.
Kanagy James M
Kinzig Charles M
SmithKline Beecham p.l.c.
Swartz Rodney P.
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