Method and apparatus for washing magnetically responsive particles
First Claim
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1. A method of washing magnetically responsive particles, the method comprising:
- flowing liquid through a flow chamber that includes magnetically responsive particles captured in a first zone by a first magnetic field;
dissipating the first magnetic field; and
agitating the magnetically responsive particles.
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Abstract
The invention features apparati and methods for washing magnetically responsive particles such as paramagnetic beads. One exemplary apparatus includes a flow chamber having an inlet and an outlet; and at least a first magnetic field inducer. The apparatus is configured such that a first magnetic field can be selectively applied in the flow chamber. The apparatus can also apply a flow of fluid to the chamber in coordination with the selective application of the magnetic field.
136 Citations
62 Claims
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1. A method of washing magnetically responsive particles, the method comprising:
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flowing liquid through a flow chamber that includes magnetically responsive particles captured in a first zone by a first magnetic field;
dissipating the first magnetic field; and
agitating the magnetically responsive particles. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 19, 20)
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15. A method comprising:
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providing a flow chamber, a first magnetic field inducer, and magnetically responsive particles disposed in the chamber, each particle physically associated with a target;
capturing the magnetically responsive particles in a first magnetic field applied by the first magnetic field inducer in the flow chamber;
flowing a solution through the flow chamber; and
displacing the magnetically responsive particles from the first zone of the flow chamber to one or more other regions of the flow chamber such that the magnetically responsive particles are retained in the flow chamber. - View Dependent Claims (16, 17, 18, 21, 22, 23, 24)
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25. A method comprising:
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a) disposing in a flow chamber magnetically responsive particles that have a target attached;
b) applying a first magnetic field to the flow chamber;
c) contacting the magnetically responsive particles to a mixture of compounds;
d) flowing liquid through the chamber; and
e) agitating the magnetically responsive particles by alternatively applying the first magnetic field and a second magnetic field. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34)
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35. A method comprising:
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providing a flow chamber that includes a mixture comprising magnetically responsive particles and members of a display library, the magnetically responsive particles having attached thereto a target compound;
washing at least some of the members from the particles by effecting one or more cycles that comprise (i) flowing a first solution through the flow chamber while the particles are captured by a magnetic field, and (ii) agitating the particles by selectively applying one or more magnetic fields. - View Dependent Claims (36, 37, 38, 39, 40, 41, 42, 43)
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44. A method comprising:
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providing a flow chamber having an inlet, an outlet and magnetically responsive particles disposed therein;
translating to a first position a frame that includes a first and second magnetic field inducer, wherein the first position locates the first magnetic field inducer such that a first magnetic field is applied to the flow chamber and the second magnetic field inducer such that a second magnetic field is not applied to the flow chamber;
flowing liquid through the flow chamber; and
translating the frame to a second position wherein the second position locates the first magnetic field inducer such that a first magnetic field is not applied to the flow chamber and the second magnetic field inducer such that a second magnetic field is applied to the flow chamber. - View Dependent Claims (45, 46)
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47. An apparatus comprising:
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a flow chamber having an inlet and an outlet; and
at least a first magnetic field inducer, wherein the apparatus is configured such that a first and second magnetic field can be selectively applied in the flow chamber.
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48. An apparatus comprising:
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a flow chamber having an inlet and an outlet and including magnetically responsive particles; and
at least a first magnetic field inducer that selectively applies a first magnetic field, wherein the magnetically responsive particles are retained in the flow chamber in the absence of the first magnetic field. - View Dependent Claims (50, 51, 52, 53, 54, 57)
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49. An apparatus comprising:
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a flow chamber having an inlet and an outlet; and
first and second magnetic field inducers, each magnetic field inducer being controllable to selectively generate apply a magnetic field in a zone of the flow chamber.
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55. An apparatus comprising:
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a support having a fitting adapted for mounting a flow chamber;
at least a first and second magnetic field inducers;
a translatable frame having attached to the magnetic field inducers; and
an actuator that translates the frame in response to a control signal, wherein the translation moves the magnetic field inducers relative to a flow chamber if mounted. - View Dependent Claims (56)
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58. A method comprising:
- forming a complex, in a vessel, that includes (a) magnetically responsive particles, (b) a target that is attached or attachable to the particles, and (c) a replicable entity that displays a heterologous protein component;
applying a magnetic field that retains the complex in the vessel;
removing fluid from the vessel;
supplying a solution that supports replication of the replicable entity to the vessel; and
replicating the first replicable entity by one or more cycles of replication.
- forming a complex, in a vessel, that includes (a) magnetically responsive particles, (b) a target that is attached or attachable to the particles, and (c) a replicable entity that displays a heterologous protein component;
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59. A method of selecting a replicable display entity, the method comprising:
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a) providing a library of replicable entities that each have a heterologous protein component that is physically attached to the respective replicable entity, wherein each protein component is a member of diverse set of different proteins;
b) contacting replicable entities of the library to a target;
c) performing one or more cycles of;
i) forming replicable entity-immobilized target complexes, each of which includes (1) a replicable entity that binds to the target by its heterologous protein component and (2) the target immobilized to a support;
ii) separating replicable entities that do not bind to the target from the replicable entity-immobilized target complexes, iii) producing copies of the replicable entities in the presence of the target, the produced copies being replicates of replicable entities that bind to the target; and
d) recovering the nucleic acid encoding the heterologous protein component of one or more produced replicable entities that bind to the target, thereby selecting a nucleic acid that encodes a binding protein for the target. - View Dependent Claims (60, 61)
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62. A method of amplifying a replicable display entity, the method comprising:
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providing a vessel that comprises a plurality of replicable entities that include a heterologous protein component and a target;
binding a first subset of the replicable entities of the plurality to the target;
separating a second subset of replicable entities of the plurality from the first subset, wherein the replicable entities of the second subset do not bind to the target; and
effecting repeated cycles of replication and solution replacement while members of the first subset are captured by the target.
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Specification