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Computer-assisted methods and apparatus for identification and characterization of biomolecules in a biological sample
| Details |
Inventors: Parekh, Rajesh Bhikhu; Amess, Robert; Bruce, James Alexander; Prime, Sally Barbara; Platt, Albert Edward; Stoney, Richard Michael;
Assignee: Oxford Glycosciences (UK) Ltd. (Abingdon, GB)
Primary Examiner: Johns; Andrew W.
Assistant Examiner:
Attorney, Agent or Firm: Pennie & Edmonds LLP
The present invention provides computer-assisted methods and apparatus for identifying, selecting and characterizing biomolecules in a biological sample. A two-dimensional array is generated by separating biomolecules present in a complex mixture. A computer-readable profile is constructed representing the identity and relative abundance of a plurality of biomolecules detected by imaging the two dimensional array. Computer-mediated comparison of profiles from multiple samples permits automated identification of subsets of biomolecules that satisfy pre-ordained criteria. Identified biomolecules can be automatically isolated from the two dimensional array by a robotic device in accordance with computer-generated instructions. A supported gel suitable for electrophoresis is provided that is bonded to a solid support such that the gel has two-dimensional spatial stability and the solid support is substantially non-interfering with respect to detection of a label, such as a fluorescent label, associated with one or more biomolecules in the gel. |
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DETAILED DESCRIPTION OF THE INVENTION The present invention provides methods and apparatus for rapidly and efficiently identifying and characterizing biomolecules, for example proteins, in a biological sample.
In one application of the invention, a biological sample is subjected to two successive separation steps.
In the first separation step, the biomolecules are separated according to one physical or chemical property so as to generate a one-dimensional array containing the biomolecules; for example, proteins are separated by isoelectric focusing along a first axis.
In the second separation step, the biomolecules in this one-dimensional array are separated according to a second physical or chemical characteristic so as to generate a two-dimensional array of separated biomolecules; for example, proteins separated by isoelectric focusing are subjected to SDS-PAGE along a second axis perpendicular to the first axis.
The separated biomolecules are stably maintained in the two-dimensional array for subsequent imaging.
The stable two-dimensional array can be stored or archived for an extended period (e.
g.
months or years) and selected biomolecules can be retrieved from the array at any desired time, based on automated computer analysis of the data derived from imaging.
The two-dimensional array is imaged with a detector to generate a computer-readable output that contains a set of x,y coordinates and a signal value for each detected biomolecule.
If desired, the computer-readable output can be displayed to a human operator--before or after computer-mediated analysis--as a computer-generated image on a screen or on any suitable medium.
Computer-mediated analysis of the computer-readable output is performed, resulting in a computer-readable profile that represents, for a plurality of detected biomolecules, the relative abundance of each such biomolecule and its attributes as deduced from its x,y coordinates in the two-dimensional array.
For example, a profile derived from imaging a gel containing proteins separated by isoelectric focusing followed by SDS-PAGE represents the isoelectric point (pI), apparent molecular weight (MW) and relative abundance of a plurality of detected proteins
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