PREDETERMINED CALIBRATION CURVE
Methods and compositions are disclosed for preparing a predetermined standardized calibration curve, which can be stored on a label, barcode or tag. A lyophilized product containing assay reagents for use in a bioassay and a known amount of analyte. A predetermined calibration curve associated with the lyophilized product. A predetermined calibration curve makes the preparation of a new calibration curve unnecessary. The methods and compositions are useful in bioassays such as a FRET assay, a real-time reverse transcription polymerase (RT-PCR) assay, a chemiluminescence assay, or any other bioassay that elicits a detectable response based on a change in, or appearance of, color, fluorescence, or reflectance.
1 . A method for preparing and storing or assigning a predetermined standardized curve for an assay, the method comprising:
(a) preparing a plurality of lyophilized calibrator compositions in an assay device, wherein the assay device has a plurality of wells or is a single cuvette;
(b) incubating the assay device in an analyzer;
(c) obtaining an optical signal for each of the plurality of standard calibrator compositions to generate a plurality of optical signals;
(d) preparing a standardized curve of the analyte from the plurality of optical signals; and
(e) storing or assigning the predetermined standardized curve into a label or barcode for the assay device.
2 . The method of claim 1 , wherein each calibrator composition contains a first antibody having a first epitope for an analyte; a second antibody having a second epitope for the analyte, and a known amount of analyte.
3 . The method of claim 2 , wherein a fluorophore acceptor is attached to the first antibody and a fluorophore donor is attached to the second antibody.
4 . The method of claim 1 , wherein each calibrator composition contains an antibody for the analyte and a known amount of analyte.
5 . The method of claim 4 , wherein a fluorophore acceptor is attached to the antibody and a fluorophore donor is attached to the analyte.
6 . The method of claim 1 , wherein the assay device is a member selected from the group consisting of a 96, 384 and 1536 well plate.
7 . The method of claim 1 , wherein the assay device is a cuvette.
8 . The method of claim 1 , wherein the analyte is a member selected from the group consisting of a protein, an autoantibody, a nucleic acid and a vitamin.
9 . A method for adjusting a predetermined standardized curve of an analyte for an assay, the method comprising:
(a) measuring a signal for a calibrator composition, wherein the calibrator composition comprises a known amount of the analyte within the predetermined standardized curve;
(b) obtaining a ratio of the signal for the calibrator composition; and
(c) adjusting the predetermined standardized curve according to the ratio obtained.
10 . The method of claim 9 , further comprising (d) optionally determining an unknown analyte concentration according to the adjusted predetermined standardized curve.
11 . The method of claim 9 , wherein the calibrator composition is a plurality of calibrator compositions.
12 . The method of claim 9 , wherein the predetermined output can be loaded onto a barcode, RFID tag or label.
13 . The method of claim 9 , wherein the ratio is used to normalize different analyzers.
14 . A lyophilized product, the lyophilized product containing:
a fluorophore acceptor attached to a first antibody having a first epitope for an analyte;
a fluorophore donor attached to a second antibody having a second epitope for the analyte; and
a known amount of analyte.
15 . The lyophilized product of claim 1 , wherein the lyophilized product is a bead.
16 . The lyophilized product of claim 15 , wherein the bead is a plurality of beads, wherein each bead has a different concentration of analyte.
17 . The lyophilized product of claim 1 , wherein the analyte is a member selected from the group consisting of a protein, a nucleic acid, an autoantibody and a vitamin.
18 . The lyophilized product of claim 17 , wherein the analyte is a protein.
19 . The lyophilized product of claim 17 , wherein the analyte is an autoantibody.
20 . The lyophilized product of claim 17 , wherein the analyte is a vitamin.